@article {19889, title = {Tripartite binding mode of cohesin-dockerin complexes from Ruminococcus flavefaciens involving naturally truncated dockerins}, volume = {301}, year = {2025}, note = {n/a}, month = {2025}, pages = {110325}, abstract = {

Polysaccharides in plant cell walls serve as a rich carbon and energy source, yet their structural complexity presents a barrier to efficient degradation. To address this, anaerobic microorganisms like R. flavefaciens have developed sophisticated multi-enzyme complexes known as cellulosomes, which enable the efficient breakdown of these recalcitrant polysaccharides. These complexes are assembled through high-affinity interactions between cohesin (Coh) modules in scaffoldin proteins and dockerin (Doc) modules in cellulosomal enzymes. R. flavefaciens FD-1 harbors one of the most intricate cellulosomes described to date, comprising over 200 Doc-containing proteins encoded in its genome. Despite substantial research on this cellulosome, the role of a group of truncated but functional dockerins, known as group-2 Docs, remains unclear. In this study, we present a detailed structural and binding analysis of a Coh-Doc complex involving the cohesin from the cell-anchoring scaffoldin ScaE and a group-2 Doc that bears only one of the two Ca+2-coordinating loops that characterise the canonical Docs. Our findings reveal a novel tripartite binding mechanism, in which the cohesin can simultaneously bind two distinct dockerin units in three alternative conformations. This discovery provides new insights into the modular versatility of the R. flavefaciens cellulosome and sheds light on the mechanisms that enhance its efficiency in polysaccharide degradation.

}, keywords = {Carbohydrates, cellulosome, cohesin, dockerin, protein assembly, protein complex, Protein structure}, isbn = {0021-9258}, url = {https://www.sciencedirect.com/science/article/pii/S0021925825021751}, author = {Duarte, Marlene and Ana Lu{\'\i}sa Carvalho and Ferreira, Magda C. and Caires, Beatriz and Rom{\~a}o, Maria Jo{\~a}o and Prates, Jos{\'e} A. M. and Najmudin, Shabir and Bayer, Edward A. and Fontes, Carlos MGA. and Bule, Pedro} } @article {Vilela-Alves:no5205, title = {{Structural and biochemical characterization of the M405S variant of {\i}t Desulfovibrio vulgaris} formate dehydrogenase}}, journal = {Acta Crystallographica Section F}, volume = {80}, number = {5}, year = {2024}, note = {n/a}, month = {May}, pages = {98{\textendash}106}, abstract = {

Molybdenum- or tungsten-dependent formate dehydrogenases have emerged as~significant catalysts for the chemical reduction of CO${\sb 2}$ to formate, with biotechnological applications envisaged in climate-change mitigation. The role of Met405 in the active site of {\i}t Desulfovibrio vulgaris} formate dehydrogenase AB ({\i}t Dv}FdhAB) has remained elusive. However, its proximity to the metal site and the conformational change that it undergoes between the resting and active forms suggests a functional role. In this work, the M405S variant was engineered, which allowed the active-site geometry in the absence of methionine S${\sp {$δ$}}$ interactions with the metal site to be revealed and the role of Met405 in catalysis to be probed. This variant displayed reduced activity in both formate oxidation and CO${\sb 2}$ reduction, together with an increased sensitivity to oxygen inactivation.

}, keywords = {Catalysis, CO2 reduction, Desulfovibrio vulgaris, metal-dependent formate dehydrogenases, Mo/W enzymes, X-ray crystallography}, doi = {10.1107/S2053230X24003911}, url = {https://doi.org/10.1107/S2053230X24003911}, author = {Vilela-Alves, Guilherme and Manuel, Rita Rebelo and Pedrosa, Neide and Cardoso Pereira, In{\^e}s A. and Rom{\~a}o, Maria Jo{\~a}o and Mota, Cristiano} } @inbook {18869, title = {CBMcarb-DB: interface of the three-dimensional landscape of carbohydrate-binding modules}, booktitle = {Carbohydrate Chemistry: Chemical and Biological Approaches Volume 46}, volume = {46}, year = {2024}, note = {n/a}, month = {2024/06/26}, publisher = {Royal Society of Chemistry}, organization = {Royal Society of Chemistry}, abstract = {

Carbohydrate-binding-modules (CBMs) are discrete auxiliary protein modules with a non-catalytic carbohydrate-binding function and that exhibit a great diversity of binding specificities. CBMcarb-DB is a curated database that classifies the three-dimensional structures of CBM{\textendash}carbohydrate complexes determined by single-crystal X-ray diffraction methods and solution NMR spectroscopy. We designed the database architecture and the navigation tools to query the database with the Protein Data Bank (PDB), UniProtKB, and GlyTouCan (universal glycan repository) identifiers. Special attention was devoted to describing the bound glycans using simple graphical representation and numerical format for cross-referencing to other glycosciences and functional data databases. CBMcarb-DB provides detailed information on CBMs and their bound oligosaccharides and features their interactions using several open-access applications. We also describe how the curated information provided by CBMcarb-DB can be integrated with AI algorithms of 3D structure prediction, facilitating structure{\textendash}function studies. Also in this chapter, we discuss the exciting convergence of CBMcarb-DB with the glycan array repository, which serves as a valuable resource for investigating the specific binding interactions between glycans and various biomolecular targets. The interaction of the two fields represents a significant milestone in glycosciences. CBMcarb-DB is freely available at https://cbmdb.glycopedia.eu/ and https://cbmcarb.webhost.fct.unl.pt.

}, isbn = {978-1-83767-217-2}, url = {https://doi.org/10.1039/BK9781837672844-00001}, author = {Ribeiro, D. O. and Bonnardel, F. and Palma, A. S. and Carvalho, A. L. M. and Perez, S.}, editor = {Pilar Rauter, Am{\'e}lia and Queneau, Yves and Palma, Angelina S{\'a}} } @article {18871, title = {An allosteric redox switch involved in oxygen protection in a CO2 reductase}, journal = {Nat Chem Biol}, volume = {20}, year = {2024}, note = {

n/a

}, month = {2024}, pages = {111 - 119}, abstract = {

Metal-dependent formate dehydrogenases reduce CO2 with high efficiency and selectivity, but are usually very oxygen sensitive. An exception is Desulfovibrio vulgaris W/Sec-FdhAB, which can be handled aerobically, but the basis for this oxygen tolerance was unknown. Here we show that FdhAB activity is controlled by a redox switch based on an allosteric disulfide bond. When this bond is closed, the enzyme is in an oxygen-tolerant resting state presenting almost no catalytic activity and very low formate affinity. Opening this bond triggers large conformational changes that propagate to the active site, resulting in high activity and high formate affinity, but also higher oxygen sensitivity. We present the structure of activated FdhAB and show that activity loss is associated with partial loss of the metal sulfido ligand. The redox switch mechanism is reversible in vivo and prevents enzyme reduction by physiological formate levels, conferring a fitness advantage during O2 exposure.

}, isbn = {1552-4469}, url = {https://doi.org/10.1038/s41589-023-01484-2}, author = {Ana Rita Oliveira and Mota, Cristiano and Vilela-Alves, Guilherme and Manuel, Rita Rebelo and Pedrosa, Neide and Fourmond, Vincent and Klymanska, Kateryna and L{\'e}ger, Christophe and Guigliarelli, Bruno and Rom{\~a}o, Maria Jo{\~a}o and Cardoso Pereira, In{\^e}s A.} } @article {19021, title = {Family GH157 enzyme exhibits broad linkage tolerance and a dual endo/exo- β -glucanase activity on β-glucans}, year = {2024}, note = {n/a}, month = {2024}, pages = {137402}, abstract = {

The structural and chemical diversity of β-glucans is reflected on the variety of essential biological roles tackled by these polysaccharides. This natural heterogeneity requires an elaborate assortment of enzymatic mechanisms to assemble, degrade or modify, as well as to extract their full biotechnological potential. Recent metagenomic efforts have provided an unprecedented growth in potential new biocatalysts, most of which remain unconfirmed or uncharacterized. Here we report the first biochemical and structural characterization of two bacterial β-glucanases from the recently created glycoside hydrolase family 157 (LaGH157 and BcGH157) and investigate their molecular basis for substrate hydrolysis. Structural analysis by X-ray crystallography revealed that GH157 enzymes belong to clan GH-A, possessing a (β/α)8-barrel fold catalytic domain, two β-sandwich accessory domains and two conserved catalytic glutamates residues, with relative positions compatible with a retaining mechanism of hydrolysis. Specificity screening and enzyme kinetics suggest that the enzymes prefer mixed-linkage glucans over β-1,3-glucans. Activity screening showed that both enzymes exhibit pH optimum at 6.5 and temperature optimum for LaGH157 and BcGH157 at 25 {\textdegree}C and 48 {\textdegree}C, respectively. Product analysis with HPAEC-PAD and LC-MS revealed that both enzymes are endo-1,3(4)-β-glucanases, capable of cleaving β-1,3 and β-1,4-linked glucoses, when preceded by a β-1,3 linkage. Moreover, BcGH157 needs a minimum of 4 subsites occupied for hydrolysis to occur, while LaGH157 only requires 3 subsites. Additionally, LaGH157 possesses exohydrolytic activity on β-1,3 and branching β-1,6 linkages. This unusual bifunctional endo-1,3(4)/exo-1,3{\textendash}1,6 activity constitutes an expansion on our understanding of β-glucan deconstruction, with the potential to inspire future applications.

}, keywords = {CAZYme, Endo-1,3(4)-β-glucanase, GH157, Glycoside hydrolase, β-Glucans}, isbn = {0141-8130}, url = {https://www.sciencedirect.com/science/article/pii/S0141813024082114}, author = {Caseiro, Catarina and McGregor, Nicholas G.S. and Alves, Victor Diogo and Ana Lu{\'\i}sa Carvalho and Rom{\~a}o, Maria Jo{\~a}o and Davies, Gideon J. and Fontes, Carlos M. G. A. and Bule, Pedro} } @article {18870, title = {Substrate-dependent oxidative inactivation of a W-dependent formate dehydrogenase involving selenocysteine displacement}, journal = {Chemical Science}, year = {2024}, note = {n/a}, month = {2024}, pages = { - }, publisher = {The Royal Society of Chemistry}, abstract = {

Metal-dependent formate dehydrogenases are very promising targets for enzyme optimization and design of bio-inspired catalysts for CO2 reduction, towards innovative strategies for climate change mitigation. For effective application of these enzymes, the catalytic mechanism must be better understood, and the molecular determinants clarified. Despite numerous studies, several doubts persist, namely regarding the role played by the possible dissociation of the SeCys ligand from the Mo/W active site. Additionally, the oxygen sensitivity of these enzymes must also be understood as it poses an important obstacle for biotechnological applications. Here we present a combined biochemical, spectroscopic, and structural characterization of Desulfovibrio vulgaris FdhAB (DvFdhAB) when exposed to oxygen in the presence of a substrate (formate or CO2). This study reveals that O2 inactivation is promoted by the presence of either substrate and involves forming a different species in the active site, captured in the crystal structures, where the SeCys ligand is displaced from tungsten coordination and replaced by a dioxygen or peroxide molecule. This form was reproducibly obtained and supports the conclusion that, although W-DvFdhAB can catalyse the oxidation of formate in the presence of oxygen for some minutes, it gets irreversibly inactivated after prolonged O2 exposure in the presence of either substrate.

}, isbn = {2041-6520}, url = {http://dx.doi.org/10.1039/D4SC02394C}, author = {Vilela-Alves, Guilherme and Manuel, Rita Rebelo and Viegas, Aldino and Philippe Carpentier and Biaso, Fr{\'e}d{\'e}ric and Guigliarelli, Bruno and Pereira, In{\^e}s Cardoso and Rom{\~a}o, Maria Jo{\~a}o and Mota, Cristiano} } @article {18758, title = {Arsenite oxidase in complex with antimonite and arsenite oxyanions: Insights into the catalytic mechanism}, journal = {Journal of Biological ChemistryJournal of Biological Chemistry}, volume = {299}, year = {2023}, note = {

doi: 10.1016/j.jbc.2023.105036

}, month = {2023}, publisher = {Elsevier}, abstract = {

Arsenic contamination of groundwater is among one of the biggest health threats affecting millions of people in the world. There is an urgent need for efficient arsenic biosensors where the use of arsenic metabolizing enzymes can be explored. In this work, we have solved four crystal structures of arsenite oxidase (Aio) in complex with arsenic and antimony oxyanions and the structures determined correspond to intermediate states of the enzymatic mechanism. These structural data were complemented with density-functional theory calculations providing a unique view of the molybdenum active site at different time points that, together with mutagenesis data, enabled to clarify the enzymatic mechanism and the molecular determinants for the oxidation of As(III) to the less toxic As(V) species.Arsenic contamination of groundwater is among one of the biggest health threats affecting millions of people in the world. There is an urgent need for efficient arsenic biosensors where the use of arsenic metabolizing enzymes can be explored. In this work, we have solved four crystal structures of arsenite oxidase (Aio) in complex with arsenic and antimony oxyanions and the structures determined correspond to intermediate states of the enzymatic mechanism. These structural data were complemented with density-functional theory calculations providing a unique view of the molybdenum active site at different time points that, together with mutagenesis data, enabled to clarify the enzymatic mechanism and the molecular determinants for the oxidation of As(III) to the less toxic As(V) species.

}, isbn = {0021-9258}, url = {https://doi.org/10.1016/j.jbc.2023.105036}, author = {Engrola, Filipa and Correia, M{\'a}rcia A. S. and Cameron Watson and Rom{\~a}o, Carlos C. and Veiros, Luis F. and Rom{\~a}o, Maria Jo{\~a}o and Santos-Silva, Teresa and Joanne M. Santini} } @article {18532, title = {Elucidating the concentration-dependent effects of thiocyanate binding to carbonic anhydrase}, volume = {244}, year = {2023}, note = {n/a}, month = {2023}, pages = {112222}, abstract = {

Many proteins naturally carry metal centers, with a large share of them being in the active sites of several enzymes. Paramagnetic effects are a powerful source of structural information and, therefore, if the native metal is paramagnetic, or it can be functionally substituted with a paramagnetic one, paramagnetic effects can be used to study the metal sites, as well as the overall structure of the protein. One notable example is cobalt(II) substitution for zinc(II) in carbonic anhydrase. In this manuscript we investigate the effects of sodium thiocyanate on the chemical environment of the metal ion of the human carbonic anhydrase II. The electron paramagnetic resonance (EPR) titration of the cobalt(II) protein with thiocyanate shows that the EPR spectrum changes from A-type to C-type on passing from 1:1 to 1:1000-fold ligand excess. This indicates the occurrence of a change in the electronic structure, which may reflect a sizable change in the metal coordination environment in turn caused by a modification of the frozen solvent glass. However, paramagnetic nuclear magnetic resonance (NMR) data indicate that the metal coordination cage remains unperturbed even in 1:1000-fold ligand excess. This result proves that the C-type EPR spectrum observed at large ligand concentration should be ascribed to the low temperature at which EPR measurements are performed, which impacts on the structure of the protein when it is destabilized by a high concentration of a chaotropic agent.

}, keywords = {EPR, Human carbonic anhydrase II, NMR, Paramagnetism, Sodium thiocyanate, Structural biology}, isbn = {0162-0134}, url = {https://www.sciencedirect.com/science/article/pii/S0162013423001046}, author = {Silva, Jos{\'e} Malanho and Cerofolini, Linda and Ana Lu{\'\i}sa Carvalho and Ravera, Enrico and Fragai, Marco and Parigi, Giacomo and Macedo, Anjos L. and Geraldes, Carlos F. G. C. and Luchinat, Claudio} } @article {18759, title = {Integrated approaches for the separation and purification of recombinant HPV16 E6 protein from Escherichia coli crude extracts}, volume = {315}, year = {2023}, note = {n/a}, month = {2023}, pages = {123647}, abstract = {

Human papillomavirus (HPV) is a sexually transmissible virus responsible for 5\% of global human cancers and associated with 99\% of cervical cancer cases. The oncogenic potential of high-risk HPVs is mainly related to the E6 and E7 oncoproteins, which are responsible, at least in part, for inactivating the p53 and pRb tumor suppressor proteins. Due to the critical role of the E6 protein in malignant tumorigenesis, it is widely recognized as a therapeutic target for anti-HPV drug development. Nevertheless, it is required to obtain large amounts of protein with high purity to perform biointeraction studies with the potential inhibitor drugs. In this work, recombinant dual-tagged E6 protein (His6-MBP-E6) was expressed from Escherichia coli (E. coli) cultures and successfully extracted by sonication/ice cycles. Affinity chromatography using MBPtrap columns allowed 85~{\textpm}~5\% protein recovery with the elimination of major host heterologous proteins in a single fraction. Subsequently, a polishing step was studied by applying anionic exchange (QSepharose), size exclusion (Superdex), or immobilized-metal affinity chromatography (HisTrap). The combination of affinity chromatography with size exclusion or two affinity chromatography techniques allowed us to obtain 82~{\textpm}~2\% and 94~{\textpm}~3\%, of highly pure His6-MBP-E6, respectively. Also, the secondary structure of His6-MBP-E6 is preserved in both purification strategies, as appraised by circular dichroism and western-blot studies. Thermal shift assay confirmed the CD results and suggested potential additives for protein stabilization. Altogether, the reproducible strategies established for the purification of His6-MBP-E6 protein could be successfully applied to later perform biointeraction studies and structural characterization of protein{\textendash}ligand complexes.

}, keywords = {Affinity purification, Chromatography, E6 protein, HPV, Protein solubility, Protein stability}, isbn = {1383-5866}, url = {https://www.sciencedirect.com/science/article/pii/S1383586623005555}, author = {Gomes, D. and Correia, M. A. S. and Rom{\~a}o, M.J. and Passarinha, L.A. and Sousa, A.} } @article {17953, title = {The structure of a Bacteroides thetaiotamicron carbohydrate-binding module provides new insight into the recognition of complex pectic polysaccharides by the human microbiome}, year = {2023}, note = {n/a}, month = {2023}, pages = {100084}, abstract = {

TheBacteroides thetaiotaomicronhas developed a consortium of enzymes capable of overcoming steric constraints and degrading, in a sequential manner, the complex rhamnogalacturonan II (RG-II) polysaccharide. BT0996 protein acts in the initial stages of the RGII depolymerisation, where its two catalytic modules remove the terminal monosaccharides from RG-II side chains A and B. BT0996 is modular and has three putative carbohydrate-binding modules (CBMs) for which the roles in the RG-II degradation are unknown. Here, we present the characterisation of themoduleat the C-terminal domain, which we designated BT0996C. The high-resolution structure obtained by X-ray crystallography reveals that the protein displays a typical β-sandwich fold with structural similarity to CBMs assigned to families 6 and 35. The distinctive features are: 1) the presence of several charged residues at the BT0996-C surface creating a large, broad positive lysine-rich patch that encompasses the putative binding site; and 2) the absence of the highly conserved binding-site signatures observed in CBMs from families 6 and 35, such as region A tryptophan and region C asparagine. These findings hint at a binding mode of BT0996-C not yet observed in its homologues. In line with this, carbohydrate microarrays and microscale thermophoresis show the ability of BT0996-C to bind α1-4-linked polygalacturonic acid, and that electrostatic interactions are essential for the recognition of the anionic polysaccharide. The results support the hypothesis that BT0996-C may have evolved to potentiate the action of BT0996 catalytic modules on the complex structure of RG-II by binding to the polygalacturonic acid backbone sequence.

}, keywords = {Carbohydrate Binding Module, Carbohydrates, Human Gut Microbiota, Rhamnogalacturonan II}, isbn = {2590-1524}, url = {https://www.sciencedirect.com/science/article/pii/S2590152422000253}, author = {Trov{\~a}o, Filipa and Correia, Viviana G. and Louren{\c c}o, Frederico M. and Ribeiro, Diana O. and Ana Lu{\'\i}sa Carvalho and Palma, Angelina S. and Pinheiro, Benedita A.} } @article {17991, title = {Structure-function studies can improve binding affinity of cohesin-dockerin interactions for multi-protein assemblies}, volume = {224}, year = {2023}, note = {n/a}, month = {2023}, pages = {55 - 67}, abstract = {

The cellulosome is an elaborate multi-enzyme structure secreted by many anaerobic microorganisms for the efficient degradation of lignocellulosic substrates. It is composed of multiple catalytic and non-catalytic components that are assembled through high-affinity protein-protein interactions between the enzyme-borne dockerin (Doc) modules and the repeated cohesin (Coh) modules present in primary scaffoldins. In some cellulosomes, primary scaffoldins can interact with adaptor and cell-anchoring scaffoldins to create structures of increasing complexity. The cellulosomal system of the ruminal bacterium, Ruminococcus flavefaciens, is one of the most intricate described to date. An unprecedent number of different Doc specificities results in an elaborate architecture, assembled exclusively through single-binding-mode type-III Coh-Doc interactions. However, a set of type-III Docs exhibits certain features associated with the classic dual-binding mode Coh-Doc interaction. Here, the structure of the adaptor scaffoldin-borne ScaH Doc in complex with the Coh from anchoring scaffoldin ScaE is described. This complex, unlike previously described type-III interactions in R. flavefaciens, was found to interact in a dual-binding mode. The key residues determining Coh recognition were also identified. This information was used to perform structure-informed protein engineering to change the electrostatic profile of the binding surface and to improve the affinity between the two modules. The results show that the nature of the residues in the ligand-binding surface plays a major role in Coh recognition and that Coh-Doc affinity can be manipulated through rational design, a key feature for the creation of designer cellulosomes or other affinity-based technologies using tailored Coh-Doc interactions.

}, keywords = {Biomass degradation, Carbohydrates, cellulosome, cohesin, dockerin, protein complex}, isbn = {0141-8130}, url = {https://www.sciencedirect.com/science/article/pii/S0141813022023480}, author = {Duarte, Marlene and Alves, Victor D. and Correia, M{\'a}rcia and Caseiro, Catarina and Ferreira, Lu{\'\i}s M. A. and Rom{\~a}o, Maria Jo{\~a}o and Ana Lu{\'\i}sa Carvalho and Najmudin, Shabir and Bayer, Edward A. and Fontes, Carlos M. G. A. and Bule, Pedro} } @article {doi:10.1128/iai.00405-22, title = {The Chlamydia trachomatis IncM Protein Interferes with Host Cell Cytokinesis, Centrosome Positioning, and Golgi Distribution and Contributes to the Stability of the Pathogen-Containing Vacuole}, journal = {Infection and Immunity}, volume = {91}, number = {4}, year = {2023}, note = {n/a}, pages = {e00405-22}, abstract = {

Chlamydia trachomatis is an obligate intracellular bacterial pathogen that causes ocular and urogenital infections in humans. The ability of C. trachomatis to grow intracellularly in a pathogen-containing vacuole (known as an inclusion) depends on chlamydial effector proteins transported into the host cell by a type III secretion system. Chlamydia trachomatis is an obligate intracellular bacterial pathogen that causes ocular and urogenital infections in humans. The ability of C. trachomatis to grow intracellularly in a pathogen-containing vacuole (known as an inclusion) depends on chlamydial effector proteins transported into the host cell by a type III secretion system. Among these effectors, several inclusion membrane proteins (Incs) insert in the vacuolar membrane. Here, we show that human cell lines infected by a C. trachomatis strain deficient for Inc CT288/CTL0540 (renamed IncM) displayed less multinucleation than when infected by IncM-producing strains (wild type or complemented). This indicated that IncM is involved in the ability of Chlamydia to inhibit host cell cytokinesis. The capacity of IncM to induce multinucleation in infected cells was shown to be conserved among its chlamydial homologues and appeared to require its two larger regions predicted to be exposed to the host cell cytosol. C. trachomatis-infected cells also displayed IncM-dependent defects in centrosome positioning, Golgi distribution around the inclusion, and morphology and stability of the inclusion. The altered morphology of inclusions containing IncM-deficient C. trachomatis was further affected by depolymerization of host cell microtubules. This was not observed after depolymerization of microfilaments, and inclusions containing wild-type C. trachomatis did not alter their morphology upon depolymerization of microtubules. Overall, these findings suggest that IncM may exert its effector function by acting directly or indirectly on host cell microtubules.

}, doi = {10.1128/iai.00405-22}, url = {https://journals.asm.org/doi/abs/10.1128/iai.00405-22}, author = {Maria Pequito Lu{\'\i}s and In{\^e}s Serrano Pereira and Joana N. Bugalh{\~a}o and Catarina N. Sim{\~o}es and Mota, Cristiano and Rom{\~a}o, Maria Jo{\~a}o and Mota, Lu{\'\i}s Jaime} } @article {10.3389/fchem.2023.1267563, title = {Hierarchical self-assembly of a reflectin-derived peptide}, journal = {Frontiers in Chemistry}, volume = {11}, year = {2023}, note = {n/a}, abstract = {

Reflectins are a family of intrinsically disordered proteins involved in cephalopod camouflage, making them an interesting source for bioinspired optical materials. Understanding reflectin assembly into higher-order structures by standard biophysical methods enables the rational design of new materials, but it is difficult due to their low solubility. To address this challenge, we aim to understand the molecular self-assembly mechanism of reflectin{\textquoteright}s basic unit{\textemdash}the protopeptide sequence YMDMSGYQ{\textemdash}as a means to understand reflectin{\textquoteright}s assembly phenomena. Protopeptide self-assembly was triggered by different environmental cues, yielding supramolecular hydrogels, and characterized by experimental and theoretical methods. Protopeptide films were also prepared to assess optical properties. Our results support the hypothesis for the protopeptide aggregation model at an atomistic level, led by hydrophilic and hydrophobic interactions mediated by tyrosine residues. Protopeptide-derived films were optically active, presenting diffuse reflectance in the visible region of the light spectrum. Hence, these results contribute to a better understanding of the protopeptide structural assembly, crucial for the design of peptide- and reflectin-based functional materials.

}, issn = {2296-2646}, doi = {10.3389/fchem.2023.1267563}, url = {https://www.frontiersin.org/articles/10.3389/fchem.2023.1267563}, author = {Dias, Ana Margarida Gon{\c c}alves Carvalho and Moreira, In{\^e}s Pimentel and Iana Lychko and Lopes Soares, C{\'a}tia and Nurrito, Arianna and Moura Barbosa, Arm{\'e}nio Jorge and Lutz-Bueno, Viviane and Mezzenga, Raffaele and Ana Lu{\'\i}sa Carvalho and Pina, Ana Sofia and Roque, Ana Cec{\'\i}lia Afonso} } @article {ijms24076246, title = {Structural Characterization of Neisseria gonorrhoeae Bacterial Peroxidase\—Insights into the Catalytic Cycle of Bacterial Peroxidases}, journal = {International Journal of Molecular Sciences}, volume = {24}, number = {7}, year = {2023}, note = {n/a}, abstract = {

Neisseria gonorrhoeae is an obligate human pathogenic bacterium responsible for gonorrhea, a sexually transmitted disease. The bacterial peroxidase, an enzyme present in the periplasm of this bacterium, detoxifies the cells against hydrogen peroxide and constitutes one of the primary defenses against exogenous and endogenous oxidative stress in this organism. The 38 kDa heterologously produced bacterial peroxidase was crystallized in the mixed-valence state, the active state, at pH 6.0, and the crystals were soaked with azide, producing the first azide-inhibited structure of this family of enzymes. The enzyme binds exogenous ligands such as cyanide and azide, which also inhibit the catalytic activity by coordinating the P heme iron, the active site, and competing with its substrate, hydrogen peroxide. The inhibition constants were estimated to be 0.4 \± 0.1 \µM and 41 \± 5 mM for cyanide and azide, respectively. Imidazole also binds and inhibits the enzyme in a more complex mechanism by binding to P and E hemes, which changes the reduction potential of the latest heme. Based on the structures now reported, the catalytic cycle of bacterial peroxidases is revisited. The inhibition studies and the crystal structure of the inhibited enzyme comprise the first platform to search and develop inhibitors that target this enzyme as a possible new strategy against N. gonorrhoeae.

}, issn = {1422-0067}, doi = {10.3390/ijms24076246}, url = {https://www.mdpi.com/1422-0067/24/7/6246}, author = {N{\'o}brega, Cl{\'a}udia S. and Ana Lu{\'\i}sa Carvalho and Rom{\~a}o, Maria Jo{\~a}o and Pauleta, Sofia R.} } @article {ijms24010476, title = {Tracking W-Formate Dehydrogenase Structural Changes During Catalysis and Enzyme Reoxidation}, journal = {International Journal of Molecular Sciences}, volume = {24}, number = {1}, year = {2023}, note = {n/a}, abstract = {

Metal-dependent formate dehydrogenases (Fdh) catalyze the reversible conversion of CO2 to formate, with unrivalled efficiency and selectivity. However, the key catalytic aspects of these enzymes remain unknown, preventing us from fully benefiting from their capabilities in terms of biotechnological applications. Here, we report a time-resolved characterization by X-ray crystallography of the Desulfovibrio vulgaris Hildenborough SeCys/W-Fdh during formate oxidation. The results allowed us to model five different intermediate structures and to chronologically map the changes occurring during enzyme reduction. Formate molecules were assigned for the first time to populate the catalytic pocket of a Fdh. Finally, the redox reversibility of DvFdhAB in crystals was confirmed by reduction and reoxidation structural studies.

}, issn = {1422-0067}, doi = {10.3390/ijms24010476}, url = {https://www.mdpi.com/1422-0067/24/1/476}, author = {Vilela-Alves, Guilherme and Manuel, Rita Rebelo and Ana Rita Oliveira and Pereira, In{\^e}s Cardoso and Rom{\~a}o, Maria Jo{\~a}o and Mota, Cristiano} } @inbook {18752, title = {The W/SeCys-FdhAB formate dehydrogenase from Desulfovibrio vulgaris Hildenborough}, booktitle = {Encyclopedia of Inorganic and Bioinorganic Chemistry}, year = {2022}, note = {

Major Reference Works

}, month = {2022/06/10}, pages = {1-12}, abstract = {

Abstract The W/SeCys-FdhAB formate dehydrogenase from Desulfovibrio vulgaris Hildenborough is a dimeric periplasmic enzyme that catalyzes the reversible oxidation of formate and reduction of CO2. It belongs to the group of metal-dependent FDHs, with a tungsten at the active site coordinated by two pyranopterin guanine dinucleotides, a selenocysteine, and one labile sulfur atom. FdhAB has a remarkably high activity and unusual tolerance to oxygen, making it an ideal model system to study biological CO2 reduction.

}, keywords = {CO2 reduction, formate dehydrogenase, pyranopterin, Selenocysteine, tungsten}, isbn = {9781119951438}, url = {https://doi.org/10.1002/9781119951438.eibc2812}, author = {Oliveira, Ana R. and Mota, Cristiano and Rom{\~a}o, Maria J. and Pereira, In{\^e}s A. C.} } @article {18753, title = {Spectroscopic and Structural Characterization of Reduced Desulfovibrio vulgaris Hildenborough W-FdhAB Reveals Stable Metal Coordination during Catalysis}, journal = {ACS Chemical BiologyACS Chemical Biology}, volume = {17}, year = {2022}, note = {

doi: 10.1021/acschembio.2c00336

}, month = {2022}, pages = {1901 - 1909}, publisher = {American Chemical Society}, abstract = {

Metal-dependent formate dehydrogenases are important enzymes due to their activity of CO2 reduction to formate. The tungsten-containing FdhAB formate dehydrogenase from Desulfovibrio vulgaris Hildenborough is a good example displaying high activity, simple composition, and a notable structural and catalytic robustness. Here, we report the first spectroscopic redox characterization of FdhAB metal centers by EPR. Titration with dithionite or formate leads to reduction of three [4Fe{\textendash}4S]1+ clusters, and full reduction requires Ti(III){\textendash}citrate. The redox potentials of the four [4Fe{\textendash}4S]1+ centers range between -250 and -530 mV. Two distinct WV signals were detected, WDV and WFV, which differ in only the g2-value. This difference can be explained by small variations in the twist angle of the two pyranopterins, as determined through DFT calculations of model compounds. The redox potential of WVI/V was determined to be -370 mV when reduced by dithionite and -340 mV when reduced by formate. The crystal structure of dithionite-reduced FdhAB was determined at high resolution (1.5 {\r A}), revealing the same structural alterations as reported for the formate-reduced structure. These results corroborate a stable six-ligand W coordination in the catalytic intermediate WV state of FdhAB.Metal-dependent formate dehydrogenases are important enzymes due to their activity of CO2 reduction to formate. The tungsten-containing FdhAB formate dehydrogenase from Desulfovibrio vulgaris Hildenborough is a good example displaying high activity, simple composition, and a notable structural and catalytic robustness. Here, we report the first spectroscopic redox characterization of FdhAB metal centers by EPR. Titration with dithionite or formate leads to reduction of three [4Fe{\textendash}4S]1+ clusters, and full reduction requires Ti(III){\textendash}citrate. The redox potentials of the four [4Fe{\textendash}4S]1+ centers range between -250 and -530 mV. Two distinct WV signals were detected, WDV and WFV, which differ in only the g2-value. This difference can be explained by small variations in the twist angle of the two pyranopterins, as determined through DFT calculations of model compounds. The redox potential of WVI/V was determined to be -370 mV when reduced by dithionite and -340 mV when reduced by formate. The crystal structure of dithionite-reduced FdhAB was determined at high resolution (1.5 {\r A}), revealing the same structural alterations as reported for the formate-reduced structure. These results corroborate a stable six-ligand W coordination in the catalytic intermediate WV state of FdhAB.

}, isbn = {1554-8929}, url = {https://doi.org/10.1021/acschembio.2c00336}, author = {Ana Rita Oliveira and Mota, Cristiano and Klymanska, Kateryna and Biaso, Fr{\'e}d{\'e}ric and Rom{\~a}o, Maria Jo{\~a}o and Guigliarelli, Bruno and Pereira, In{\^e}s Cardoso} } @article {ijms23137264, title = {Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation}, journal = {International Journal of Molecular Sciences}, volume = {23}, number = {13}, year = {2022}, note = {n/a}, abstract = {

Membrane-bound catechol-O-methyltransferase (MBCOMT), present in the brain and involved in the main pathway of the catechol neurotransmitter deactivation, is linked to several types of human dementia, which are relevant pharmacological targets for new potent and nontoxic inhibitors that have been developed, particularly for Parkinson\’s disease treatment. However, the inexistence of an MBCOMT 3D-structure presents a blockage in new drugs\’ design and clinical studies due to its instability. The enzyme has a clear tendency to lose its biological activity in a short period of time. To avoid the enzyme sequestering into a non-native state during the downstream processing, a multi-component buffer plays a major role, with the addition of additives such as cysteine, glycerol, and trehalose showing promising results towards minimizing hMBCOMT damage and enhancing its stability. In addition, ionic liquids, due to their virtually unlimited choices for cation/anion paring, are potential protein stabilizers for the process and storage buffers. Screening experiments were designed to evaluate the effect of distinct cation/anion ILs interaction in hMBCOMT enzymatic activity. The ionic liquids: choline glutamate [Ch][Glu], choline dihydrogen phosphate ([Ch][DHP]), choline chloride ([Ch]Cl), 1- dodecyl-3-methylimidazolium chloride ([C12mim]Cl), and 1-butyl-3-methylimidazolium chloride ([C4mim]Cl) were supplemented to hMBCOMT lysates in a concentration from 5 to 500 mM. A major potential stabilizing effect was obtained using [Ch][DHP] (10 and 50 mM). From the DoE 146\% of hMBCOMT activity recovery was obtained with [Ch][DHP] optimal conditions (7.5 mM) at \−80 \°C during 32.4 h. These results are of crucial importance for further drug development once the enzyme can be stabilized for longer periods of time.

}, issn = {1422-0067}, doi = {10.3390/ijms23137264}, url = {https://www.mdpi.com/1422-0067/23/13/7264}, author = {Gon{\c c}alves, Ana M. and Sousa, {\^A}ngela and Pedro, Augusto Q. and Rom{\~a}o, Maria J. and Jo{\~a}o A. Queiroz and Gallardo, Eug{\'e}nia and Passarinha, Lu{\'\i}s A.} } @article {MOREIRA2022100290, title = {Synergy between silk fibroin and ionic liquids for active gas-sensing materials}, journal = {Materials Today Bio}, year = {2022}, note = {n/a}, pages = {100290}, abstract = {

Silk fibroin is a biobased material with excellent biocompatibility and mechanical properties, but its use in bioelectronics is hampered by the difficult dissolution and low intrinsic conductivity. Some ionic liquids are known to dissolve fibroin but removed after fibroin processing. However, ionic liquids and fibroin can cooperatively give rise to functional materials, and there are untapped opportunities in this combination. The dissolution of fibroin, followed by gelation, in designer ionic liquids from the imidazolium chloride family with varied alkyl chain lengths (2{\textendash}10 carbons) is shown here. The alkyl chain length of the anion has a large impact on fibroin secondary structure which adopts unconventional arrangements, yielding robust gels with distinct hierarchical organization. Furthermore, and due to their remarkable air-stability and ionic conductivity, fibroin ionogels are exploited as active electrical gas sensors in an electronic nose revealing the unravelled possibilities of fibroin in soft and flexible electronics.

}, keywords = {Bioelectronics, gas sensing, Ionic conductivity, ionic liquids, Physical ionogels, Silk fibroin}, issn = {2590-0064}, doi = {https://doi.org/10.1016/j.mtbio.2022.100290}, url = {https://www.sciencedirect.com/science/article/pii/S2590006422000886}, author = {In{\^e}s P. Moreira and Carina Esteves and Susana I C J Palma and Ramou, Efthymia and Ana L.M. Carvalho and Roque, Ana C A} } @article {https://doi.org/10.1002/adma.202107205, title = {Tackling Humidity with Designer Ionic Liquid-Based Gas Sensing Soft Materials}, journal = {Advanced Materials}, volume = {34}, number = {8}, year = {2022}, note = {n/a}, pages = {2107205}, abstract = {

Abstract Relative humidity is simultaneously a sensing target and a contaminant in gas and volatile organic compound (VOC) sensing systems, where strategies to control humidity interference are required. An unmet challenge is the creation of gas-sensitive materials where the response to humidity is controlled by the material itself. Here, humidity effects are controlled through the design of gelatin formulations in ionic liquids without and with liquid crystals as electrical and optical sensors, respectively. In this design, the anions [DCA]- and [Cl]- of room temperature ionic liquids from the 1-butyl-3-methylimidazolium family tailor the response to humidity and, subsequently, sensing of VOCs in dry and humid conditions. Due to the combined effect of the materials formulations and sensing mechanisms, changing the anion from [DCA]- to the much more hygroscopic [Cl]-, leads to stronger electrical responses and much weaker optical responses to humidity. Thus, either humidity sensors or humidity-tolerant VOC sensors that do not require sample preconditioning or signal processing to correct humidity impact are obtained. With the wide spread of 3D- and 4D-printing and intelligent devices, the monitoring and tuning of humidity in sustainable biobased materials offers excellent opportunities in e-nose sensing arrays and wearable devices compatible with operation at room conditions.

}, keywords = {anion-tunability, gas sensing, gelatin, humidity, ionogels, liquid crystals, methylimidazolium ionic liquids}, doi = {https://doi.org/10.1002/adma.202107205}, url = {https://onlinelibrary.wiley.com/doi/abs/10.1002/adma.202107205}, author = {Carina Esteves and Susana I C J Palma and Henrique M.A. Costa and Cl{\'a}udia Alves and Gon{\c c}alo M.C. Santos and Ramou, Efthymia and Ana Lu{\'\i}sa Carvalho and Vitor Alves and Roque, Ana C A} } @article {16716, title = {Combined in silico and in vitro studies to identify novel antidiabetic flavonoids targeting glycogen phosphorylase}, volume = {108}, year = {2021}, note = {n/a}, month = {2021}, pages = {104552}, abstract = {

Novel pharmacological strategies for the treatment of diabetic patients are now focusing on inhibiting glycogenolysis steps. In this regard, glycogen phosphorylase (GP) is a validated target for the discovery of innovative antihyperglycemic molecules. Natural products, and in particular flavonoids, have been reported as potent inhibitors of GP at the cellular level. Herein, free-energy calculations and microscale thermophoresis approaches were performed to get an in-depth assessment of the binding affinities and elucidate intermolecular interactions of several flavonoids at the inhibitor site of GP. To our knowledge, this is the first study indicating genistein, 8-prenylgenistein, apigenin, 8-prenylapigenin, 8-prenylnaringenin, galangin and valoneic acid dilactone as natural molecules with high inhibitory potency toward GP. We identified: i) the residues Phe285, Tyr613, Glu382 and/or Arg770 as the most relevant for the binding of the best flavonoids to the inhibitor site of GP, and ii) the 5-OH, 7-OH, 8-prenyl substitutions in ring A and the 4'-OH insertion in ring B to favor flavonoid binding at this site. Our results are invaluable to plan further structural modifications through organic synthesis approaches and develop more effective pharmaceuticals for Type 2 Diabetes treatment, and serve as the starting point for the exploration of food products for therapeutic usage, as well as for the development of novel bio-functional food and dietary supplements/herbal medicines.

}, keywords = {Biophysical chemistry, Enzyme inhibition, Free-energy calculations, Microscale thermophoresis, Prenylflavonoids, Type 2 diabetes}, isbn = {0045-2068}, url = {https://www.sciencedirect.com/science/article/pii/S0045206820318502}, author = {Br{\'a}s, Nat{\'e}rcia F. and Neves, Rui P.P. and Lopes, Filipa A.A. and Correia, M{\'a}rcia A. S. and Palma, Angelina S. and Sousa, S{\'e}rgio F. and Ramos, Maria J.} } @article {16972, title = {Interrogating the Inhibition Mechanisms of Human Aldehyde Oxidase by X-ray Crystallography and NMR Spectroscopy: The Raloxifene Case}, journal = {Journal of Medicinal ChemistryJournal of Medicinal Chemistry}, year = {2021}, note = {

doi: 10.1021/acs.jmedchem.1c01125

}, month = {2021}, publisher = {American Chemical Society}, abstract = {

Human aldehyde oxidase (hAOX1) is mainly present in the liver and has an emerging role in drug metabolism, since it accepts a wide range of molecules as substrates and inhibitors. Herein, we employed an integrative approach by combining NMR, X-ray crystallography, and enzyme inhibition kinetics to understand the inhibition modes of three hAOX1 inhibitors{\textemdash}thioridazine, benzamidine, and raloxifene. These integrative data indicate that thioridazine is a noncompetitive inhibitor, while benzamidine presents a mixed type of inhibition. Additionally, we describe the first crystal structure of hAOX1 in complex with raloxifene. Raloxifene binds tightly at the entrance of the substrate tunnel, stabilizing the flexible entrance gates and elucidating an unusual substrate-dependent mechanism of inhibition with potential impact on drug{\textendash}drug interactions. This study can be considered as a proof-of-concept for an efficient experimental screening of prospective substrates and inhibitors of hAOX1 relevant in drug discovery.Human aldehyde oxidase (hAOX1) is mainly present in the liver and has an emerging role in drug metabolism, since it accepts a wide range of molecules as substrates and inhibitors. Herein, we employed an integrative approach by combining NMR, X-ray crystallography, and enzyme inhibition kinetics to understand the inhibition modes of three hAOX1 inhibitors{\textemdash}thioridazine, benzamidine, and raloxifene. These integrative data indicate that thioridazine is a noncompetitive inhibitor, while benzamidine presents a mixed type of inhibition. Additionally, we describe the first crystal structure of hAOX1 in complex with raloxifene. Raloxifene binds tightly at the entrance of the substrate tunnel, stabilizing the flexible entrance gates and elucidating an unusual substrate-dependent mechanism of inhibition with potential impact on drug{\textendash}drug interactions. This study can be considered as a proof-of-concept for an efficient experimental screening of prospective substrates and inhibitors of hAOX1 relevant in drug discovery.

}, isbn = {0022-2623}, url = {https://doi.org/10.1021/acs.jmedchem.1c01125}, author = {Mota, Cristiano and Diniz, Ana and Coelho, Catarina and Santos-Silva, Teresa and Esmaeeli, Mariam and Leimk{\"u}hler, Silke and Cabrita, Eurico J. and Marcelo, Filipa and Rom{\~a}o, Maria Jo{\~a}o} } @article {16987, title = {The SOUL family of heme-binding proteins: Structure and function 15~years later}, volume = {448}, year = {2021}, note = {n/a}, month = {2021}, pages = {214189}, abstract = {

The SOUL, or heme-binding protein HBP/SOUL, family represents a group of evolutionary conserved putative heme-binding proteins that contains a number of members in animal, plant andbacterial species. The structures of the murine form of HEBP1, or p22HBP, and the human form of HEBP2, or SOUL, have been determined in 2006 and 2011 respectively. In this work we discuss the structures of HEBP1 and HEBP2 in light of new X-ray data for heme bound murine HEBP1. The interaction between tetrapyrroles and HEBP1, initially proven to be hydrophobic in nature, was thought to also involve electrostatic interactions between heme propionate groups and positively charged amino acid side chains. However, the new X-ray structure, and results from murine HEBP1 variants and human HEBP1, confirm the hydrophobic nature of the heme-HEBP1 interaction, resulting in Kd values in the low nanomolar range, and rules out any electrostatic stabilization. Results from NMR relaxation time measurements for human HEBP1 describe a rigid globular protein with no change in motional regime upon heme binding. X-ray structures deposited in the PDB for human HEBP2 are very similar to each other and to the new heme-bound murine HEBP1 X-ray structure (backbone rmsd ca. 1~{\r A}). Results from a HSQC spectrum centred on the histidine side chain Nδ-proton region for HEBP2 confirm that HEBP2 does not bind heme via H42 as no chemical shift differences were observed upon heme addition for backbone NH and Nδ protons. A survey of the functions attributed to HEBP1 and HEBP2 over the last 20~years span a wide range of cellular pathways. Interestingly, many of them are specific to higher eukaryotes, particularly mammals and a potential link between heme release under oxidative stress and human HEBP1 is also examined using recent data. However, at the present moment, trying to relate function to the involvement of heme or tetrapyrrole binding, specifically, makes little sense with our current biological knowledge and can only be applied to HEBP1, as HEBP2 does not interact with heme. We suggest that it may not be justified to call this very small family of proteins, heme-binding proteins. The family may be more correctly called {\textquotedblleft}the SOUL family of proteins related to cellular fate{\textquotedblright} as, even though only HEBP1 binds heme tightly, both proteins may be involved in cell survival and/or proliferation.

}, keywords = {Function, HEBP1, HEBP2, NMR spectroscopy, SOUL protein, structure, X-ray crystallography}, isbn = {0010-8545}, url = {https://www.sciencedirect.com/science/article/pii/S001085452100463X}, author = {Goodfellow, Brian J. and Freire, Filipe and Ana Lu{\'\i}sa Carvalho and Aveiro, Susana S. and Charbonnier, Peggy and Moulis, Jean-Marc and Delgado, Leonildo and Gloria C. Ferreira and Rodrigues, Jo{\~a}o E. and Poussin-Courmontagne, Pierre and Birck, Catherine and McEwen, Alastair and Macedo, Anjos L.} } @article {https://doi.org/10.1002/ejic.202000868, title = {Donor Functionalized Iron(II) N-Heterocyclic Carbene Complexes in Transfer Hydrogenation Reactions}, journal = {European Journal of Inorganic Chemistry}, volume = {2021}, number = {1}, year = {2021}, note = {n/a}, pages = {22-29}, abstract = {

Two piano-stool iron(II) complexes bearing N-heterocyclic carbene ligands outfitted with acetamide- and amine-pendant arms [Cp*Fe(NHCR)(CO)I] {Cp* = η5-tetramethylcyclopentadienyl; R = CH2CONEt2 (3), (CH2)2NEt2 (4)}, have been prepared and fully characterized. Their catalytic activity in transfer hydrogenation (TH) of ketones using iPrOH as a hydrogen source and catalytic amounts of base (LiOtBu) has been explored, along with that of previously reported [CpFe(NHCR)(CO)I] {R = nBu (5), (CH2)2OH (6), Et (7), and (CH2)3OH (8)} complexes containing hydroxyl and nonfunctionalized alkyl arms. Complex 3 displayed the highest catalytic activity of the whole series 3{\textendash}8, reaching a TOF50 value of 533 h{\textendash}1. NMR monitoring of the stoichiometric reaction of 3 with LiOtBu, allowed the identification of a new species 3{\textquoteright} containing a deprotonated amidate moiety, which has been fully characterized by 1H, 13C, and 15N NMR. Finally, a green protocol for the reduction of ketones through TH using glycerol as a hydrogen source, under microwave irradiation in the presence of catalytic amounts of 3 and base has been developed.

}, keywords = {Carbene ligands, Iron, Ketones, Microwave irradiation, Reduction, Transfer hydrogenation}, doi = {https://doi.org/10.1002/ejic.202000868}, url = {https://chemistry-europe.onlinelibrary.wiley.com/doi/abs/10.1002/ejic.202000868}, author = {Lopes, Rita and Raya-Bar{\'o}n, {\'A}lvaro and Robalo, M. Paula and Vinagreiro, Carolina and Barroso, S{\'o}nia and Rom{\~a}o, Maria J. and Fern{\'a}ndez, Ignacio and Pereira, Mariette M. and Royo, Beatriz} } @article {ijms221810012, title = {Enhanced Stability of Detergent-Free Human Native STEAP1 Protein from Neoplastic Prostate Cancer Cells upon an Innovative Isolation Procedure}, journal = {International Journal of Molecular Sciences}, volume = {22}, number = {18}, year = {2021}, note = {n/a}, abstract = {

Background: The STEAP1 is a cell-surface antigen over-expressed in prostate cancer, which contributes to tumor progression and aggressiveness. However, the molecular mechanisms underlying STEAP1 and its structural determinants remain elusive. Methods: The fraction capacity of Butyl- and Octyl-Sepharose matrices on LNCaP lysates was evaluated by manipulating the ionic strength of binding and elution phases, followed by a Co-Immunoprecipitation (Co-IP) polishing. Several potential stabilizing additives were assessed, and the melting temperature (Tm) values ranked the best/worst compounds. The secondary structure of STEAP1 was identified by circular dichroism. Results: The STEAP1 was not fully captured with 1.375 M (Butyl), in contrast with interfering heterologous proteins, which were strongly retained and mostly eluted with water. This single step demonstrated higher selectivity of Butyl-Sepharose for host impurities removal from injected crude samples. Co-IP allowed recovering a purified fraction of STEAP1 and contributed to unveil potential physiologically interacting counterparts with the target. A Tm of ~55 {\textdegree}C was determined, confirming STEAP1 stability in the purification buffer. A predominant α-helical structure was identified, ensuring the protein{\textquoteright}s structural stability. Conclusions: A method for successfully isolating human STEAP1 from LNCaP cells was provided, avoiding the use of detergents to achieve stability, even outside a membrane-mimicking environment.

}, issn = {1422-0067}, doi = {10.3390/ijms221810012}, url = {https://www.mdpi.com/1422-0067/22/18/10012}, author = {Barroca-Ferreira, Jorge and Cruz-Vicente, Pedro and Santos, Marino F. A. and Rocha, Sandra M. and Santos-Silva, Teresa and Maia, Cl{\'a}udio J. and Passarinha, Lu{\'\i}s A.} } @article {D0CE01529F, title = {Magnetic particles used in a new approach for designed protein crystallization}, journal = {CrystEngComm}, volume = {23}, year = {2021}, note = {n/a}, pages = {1083-1090}, publisher = {The Royal Society of Chemistry}, abstract = {

After more than one hundred and thirty thousand protein structures determined by X-ray crystallography{,} the challenge of protein crystallization for 3D structure determination remains. In the quest for additives for efficient protein crystallization{,} inorganic materials emerge as an alternative. Magnetic particles (MPs) are versatile inorganic materials{,} easy to use{,} modify and manipulate in a wide range of biological assays. The potential of using functionalised MPs as crystallization chaperones for protein crystallization was shown in this work. MPs with distinct coatings were rationally designed to promote protein crystallization by affinity-triggered heterogeneous nucleation. Hen egg white lysozyme (HEWL) and trypsin{,} were crystallized in the presence of MPs either bare or coated with a polysaccharide (chitin) or a protein (casein){,} respectively. The addition of MPs was characterized in terms of bound protein to the MPs{,} crystal morphology{,} time-lapse of crystal emergence{,} crystallization yield fold change and crystal diffraction quality for structure determination. The MPs additives have shown to bind to the respective target protein{,} and to promote nucleation and crystal growth without compromising crystal morphology. On the other hand{,} MPs addition led to faster detectable crystal emergence and up to 13 times higher crystallization yield{,} addressing some the challenges in protein crystallization{,} the main bottleneck of macromolecular crystallography. Structure determination of the protein crystallized in the presence of MPs revealed that the structural characteristics of the protein remained unchanged{,} as shown by the superposition with PDB annotated proteins. Moreover{,} and unlike most reported cases{,} it was possible to exclude the inhibitor benzamidine during trypsin crystallisation{,} which is a remarkable result opening new prospects in enzyme engineering and drug design. Our results show that MPs coated with affinity ligands to target proteins can be used as controlled and tailor-made crystallization inducers.

}, doi = {10.1039/D0CE01529F}, url = {http://dx.doi.org/10.1039/D0CE01529F}, author = {Raquel dos Santos and Rom{\~a}o, Maria Jo{\~a}o and Roque, Ana Cec{\'\i}lia A and Ana Lu{\'\i}sa Carvalho} } @article {membranes11080549, title = {Protein Crystallization in a Microfluidic Contactor with Nafion{\textregistered}117 Membranes}, journal = {Membranes}, volume = {11}, number = {8}, year = {2021}, note = {n/a}, abstract = {

Protein crystallization still remains mostly an empirical science, as the production of crystals with the required quality for X-ray analysis is dependent on the intensive screening of the best protein crystallization and crystal{\textquoteright}s derivatization conditions. Herein, this demanding step was addressed by the development of a high-throughput and low-budget microfluidic platform consisting of an ion exchange membrane (117 Nafion{\textregistered} membrane) sandwiched between a channel layer (stripping phase compartment) and a wells layer (feed phase compartment) forming 75 independent micro-contactors. This microfluidic device allows for a simultaneous and independent screening of multiple protein crystallization and crystal derivatization conditions, using Hen Egg White Lysozyme (HEWL) as the model protein and Hg2+ as the derivatizing agent. This microdevice offers well-regulated crystallization and subsequent crystal derivatization processes based on the controlled transport of water and ions provided by the 117 Nafion{\textregistered} membrane. Diffusion coefficients of water and the derivatizing agent (Hg2+) were evaluated, showing the positive influence of the protein drop volume on the number of crystals and crystal size. This microfluidic system allowed for crystals with good structural stability and high X-ray diffraction quality and, thus, it is regarded as an efficient tool that may contribute to the enhancement of the proteins{\textquoteright} crystals structural resolution.

}, issn = {2077-0375}, doi = {10.3390/membranes11080549}, url = {https://www.mdpi.com/2077-0375/11/8/549}, author = {Polino, M. and Rho, H. S. and Pina, M. P. and Mallada, R. and Carvalho, A. L. and Rom{\~a}o, M.J. and Coelhoso, Isabel and Gardeniers, J. G. E. and Crespo, J. G. and Portugal, Carla A. M.} } @article {16943, title = {Catalytic Mechanism of Human Aldehyde Oxidase}, journal = {ACS CatalysisACS Catalysis}, volume = {10}, year = {2020}, note = {

doi: 10.1021/acscatal.0c02627

}, month = {2020}, pages = {9276 - 9286}, publisher = {American Chemical Society}, abstract = {

The mechanism of oxidation of N-heterocycle phthalazine to phthalazin-1(2H)-one and its associated free energy profile, catalyzed by human aldehyde oxidase (hAOX1), was studied in atomistic detail using QM/MM methodologies. The studied reaction was found to involve three sequential steps: (i) protonation of the substrate{\textquoteright}s N2 atom by Lys893, (ii) nucleophilic attack of the hydroxyl group of the molybdenum cofactor (Moco) to the substrate, and (iii) hydride transfer from the substrate to the sulfur atom of the Moco. The free energy profile that was calculated revealed that the rate-limiting step corresponds to hydride transfer. It was also found that Lys893 plays a relevant role in the reaction, being important not only for the anchorage of the substrate close to the Moco, but also in the catalytic reaction. The variations of the oxidation state of the molybdenum ion throughout the catalytic cycle were examined too. We found out that during the displacement of the products away from the Moco, the transfer of electrons from the catalytic site to the FAD site was proton-coupled. As a consequence, the most favorable and fastest pathway for the enzyme to complete its catalytic cycle was that with MoV and a deprotonated SH ligand of the Moco with the FAD molecule converted to its semiquinone form, FADH{\textbullet}.The mechanism of oxidation of N-heterocycle phthalazine to phthalazin-1(2H)-one and its associated free energy profile, catalyzed by human aldehyde oxidase (hAOX1), was studied in atomistic detail using QM/MM methodologies. The studied reaction was found to involve three sequential steps: (i) protonation of the substrate{\textquoteright}s N2 atom by Lys893, (ii) nucleophilic attack of the hydroxyl group of the molybdenum cofactor (Moco) to the substrate, and (iii) hydride transfer from the substrate to the sulfur atom of the Moco. The free energy profile that was calculated revealed that the rate-limiting step corresponds to hydride transfer. It was also found that Lys893 plays a relevant role in the reaction, being important not only for the anchorage of the substrate close to the Moco, but also in the catalytic reaction. The variations of the oxidation state of the molybdenum ion throughout the catalytic cycle were examined too. We found out that during the displacement of the products away from the Moco, the transfer of electrons from the catalytic site to the FAD site was proton-coupled. As a consequence, the most favorable and fastest pathway for the enzyme to complete its catalytic cycle was that with MoV and a deprotonated SH ligand of the Moco with the FAD molecule converted to its semiquinone form, FADH{\textbullet}.

}, url = {https://doi.org/10.1021/acscatal.0c02627}, author = {Ferreira, Pedro and Cerqueira, Nuno M. F. Sousa A. and Fernandes, Pedro Alexandrino and Rom{\~a}o, Maria Jo{\~a}o and Ramos, Maria Jo{\~a}o} } @article {16778, title = {Evolution, expression, and substrate specificities of aldehyde oxidase enzymes in eukaryotes}, journal = {Journal of Biological ChemistryJournal of Biological Chemistry}, volume = {295}, year = {2020}, note = {

doi: 10.1074/jbc.REV119.007741

}, month = {2020}, pages = {5377 - 5389}, publisher = {Elsevier}, abstract = {

Aldehyde oxidases (AOXs) are a small group of enzymes belonging to the larger family of molybdo-flavoenzymes, along with the well-characterized xanthine oxidoreductase. The two major types of reactions that are catalyzed by AOXs are the hydroxylation of heterocycles and the oxidation of aldehydes to their corresponding carboxylic acids. Different animal species have different complements of AOX genes. The two extremes are represented in humans and rodents; whereas the human genome contains a single active gene (AOX1), those of rodents, such as mice, are endowed with four genes (Aox1-4), clustering on the same chromosome, each encoding a functionally distinct AOX enzyme. It still remains enigmatic why some species have numerous AOX enzymes, whereas others harbor only one functional enzyme. At present, little is known about the physiological relevance of AOX enzymes in humans and their additional forms in other mammals. These enzymes are expressed in the liver and play an important role in the metabolisms of drugs and other xenobiotics. In this review, we discuss the expression, tissue-specific roles, and substrate specificities of the different mammalian AOX enzymes and highlight insights into their physiological roles.Aldehyde oxidases (AOXs) are a small group of enzymes belonging to the larger family of molybdo-flavoenzymes, along with the well-characterized xanthine oxidoreductase. The two major types of reactions that are catalyzed by AOXs are the hydroxylation of heterocycles and the oxidation of aldehydes to their corresponding carboxylic acids. Different animal species have different complements of AOX genes. The two extremes are represented in humans and rodents; whereas the human genome contains a single active gene (AOX1), those of rodents, such as mice, are endowed with four genes (Aox1-4), clustering on the same chromosome, each encoding a functionally distinct AOX enzyme. It still remains enigmatic why some species have numerous AOX enzymes, whereas others harbor only one functional enzyme. At present, little is known about the physiological relevance of AOX enzymes in humans and their additional forms in other mammals. These enzymes are expressed in the liver and play an important role in the metabolisms of drugs and other xenobiotics. In this review, we discuss the expression, tissue-specific roles, and substrate specificities of the different mammalian AOX enzymes and highlight insights into their physiological roles.

}, isbn = {0021-9258}, url = {https://doi.org/10.1074/jbc.REV119.007741}, author = {Terao, Mineko and Garattini, Enrico and Rom{\~a}o, Maria Jo{\~a}o and Leimk{\"u}hler, Silke} } @article {15667, title = {Improving the Anti-inflammatory Response via Gold Nanoparticle Vectorization of CO-Releasing Molecules}, journal = {ACS Biomaterials Science \& EngineeringACS Biomaterials Science \& Engineering}, volume = {6}, year = {2020}, note = {

doi: 10.1021/acsbiomaterials.9b01936

}, month = {2020}, pages = {1090 - 1101}, publisher = {American Chemical Society}, abstract = {

CO-releasing molecules (CORMs) have been widely studied for their anti-inflammatory, antiapoptotic, and antiproliferative effects. CORM-3 is a water-soluble Ru-based metal carbonyl complex, which metallates serum proteins and readily releases CO in biological media. In this work, we evaluated the anti-inflammatory and wound-healing effects of gold nanoparticles{\textendash}CORM-3 conjugates, AuNPs@PEG@BSA{\textperiodcentered}Ru(CO)x, exploring its use as an efficient CO carrier. Our results suggest that the nanoformulation was capable of inducing a more pronounced cell effect, at the anti-inflammatory level and a faster tissue repair, probably derived from a rapid cell uptake of the nanoformulation that results in the increase of CO inside the cell.CO-releasing molecules (CORMs) have been widely studied for their anti-inflammatory, antiapoptotic, and antiproliferative effects. CORM-3 is a water-soluble Ru-based metal carbonyl complex, which metallates serum proteins and readily releases CO in biological media. In this work, we evaluated the anti-inflammatory and wound-healing effects of gold nanoparticles{\textendash}CORM-3 conjugates, AuNPs@PEG@BSA{\textperiodcentered}Ru(CO)x, exploring its use as an efficient CO carrier. Our results suggest that the nanoformulation was capable of inducing a more pronounced cell effect, at the anti-inflammatory level and a faster tissue repair, probably derived from a rapid cell uptake of the nanoformulation that results in the increase of CO inside the cell.

}, url = {https://doi.org/10.1021/acsbiomaterials.9b01936}, author = {Alexandra R. Fernandes and Mendon{\c c}a-Martins, In{\^e}s and Santos, Marino F. A. and Lu{\'\i}s R. Raposo and Rita Mendes and Marques, Joana and Rom{\~a}o, Carlos C. and Rom{\~a}o, Maria Jo{\~a}o and Santos-Silva, Teresa and Pedro V. Baptista} } @article {15710, title = {Magnetic Precipitation: A New Platform for Protein Purification}, journal = {Biotechnology JournalBiotechnology Journal}, volume = {n/a}, year = {2020}, note = {

doi: 10.1002/biot.202000151

}, month = {2020}, pages = {2000151}, publisher = {John Wiley \& Sons, Ltd}, abstract = {

One of the trends in downstream processing comprises the use of ?anything-but-chromatography? methods to overcome the current downfalls of standard packed-bed chromatography. Precipitation and magnetic separation are two techniques already proven to accomplish protein purification from complex media, yet never used in synergy. With the aim to capture antibodies directly from crude extracts, a new approach combining precipitation and magnetic separation was developed and named as affinity magnetic precipitation. A precipitation screening, based on the Hofmeister series, and a commercial precipitation kit were tested with affinity magnetic particles to assess the best condition for antibody capture from human serum plasma and clarified cell supernatant. The best conditions were obtained when using PEG3350 as precipitant at 4{\textdegree}C for 1h, reaching 80\% purity and 50\% recovery of polyclonal antibodies from plasma, and 99\% purity with 97\% recovery yield of anti-TNFα mAb from cell supernatants. These results show that the synergetic use of precipitation and magnetic separation can represent an alternative for the efficient capture of antibodies. This article is protected by copyright. All rights reserved

}, keywords = {affinity magnetic precipitation, antibody purification, downstream, Magnetic nanoparticles, method development}, isbn = {1860-6768}, url = {https://doi.org/10.1002/biot.202000151}, author = {Raquel dos Santos and Iria, In{\^e}s and Manuel, Ana M and Leandro, Ana P and Madeira, Catarina A C and Goncalves, Joao and Ana Lu{\'\i}sa Carvalho and Roque, Ana Cec{\'\i}lia} } @article {16658, title = {Multitask ATPases (NBDs) of bacterial ABC importers type I and their interspecies exchangeability}, volume = {10}, year = {2020}, note = {n/a}, month = {2020}, pages = {19564}, abstract = {

ATP-binding cassette (ABC) type I importers are widespread in bacteria and play a crucial role in its survival and pathogenesis. They share the same modular architecture comprising two intracellular nucleotide-binding domains (NBDs), two transmembrane domains (TMDs) and a substrate-binding protein. The NBDs bind and hydrolyze ATP, thereby generating conformational changes that are coupled to the TMDs and lead to substrate translocation. A group of multitask NBDs that are able to serve as the cellular motor for multiple sugar importers was recently discovered. To understand why some ABC importers share energy-coupling components, we used the MsmX ATPase from Bacillus subtilis as a model for biological and structural studies. Here we report the first examples of functional hybrid interspecies ABC type I importers in which the NBDs could be exchanged. Furthermore, the first crystal structure of an assigned multitask NBD provides a framework to understand the molecular basis of the broader specificity of interaction with the TMDs.

}, isbn = {2045-2322}, url = {https://doi.org/10.1038/s41598-020-76444-0}, author = {Leisico, Francisco and Lia M. Godinho and Gon{\c c}alves, In{\^e}s C. and Silva, Sara P. and Carneiro, Bruno and Rom{\~a}o, Maria J. and Santos-Silva, Teresa and Isabel de S{\'a}-Nogueira} } @article {15134, title = {SLMP53-1 interacts with wild-type and mutant p53 DNA-binding domain and reactivates multiple hotspot mutations}, volume = {1864}, year = {2020}, note = {n/a}, month = {2020}, pages = {129440}, abstract = {

BackgroundHalf of human cancers harbour TP53 mutations that render p53 inactive as a tumor suppressor. As such, reactivation of mutant (mut)p53 through restoration of wild-type (wt)-like function represents one of the most promising therapeutic strategies in cancer treatment. Recently, we have reported the (S)-tryptophanol-derived oxazoloisoindolinone SLMP53-1 as a new reactivator of wt and mutp53 R280K with in vitro and in vivo p53-dependent antitumor activity. The present work aimed a mechanistic elucidation of mutp53 reactivation by SLMP53-1.
Methods and results
By cellular thermal shift assay (CETSA), it is shown that SLMP53-1 induces wt and mutp53 R280K thermal stabilization, which is indicative of intermolecular interactions with these proteins. Accordingly, in silico studies of wt and mutp53 R280K DNA-binding domain with SLMP53-1 unveiled that the compound binds at the interface of the p53 homodimer with the DNA minor groove. Additionally, using yeast and p53-null tumor cells ectopically expressing distinct highly prevalent mutp53, the ability of SLMP53-1 to reactivate multiple mutp53 is evidenced.
Conclusions
SLMP53-1 is a p53-activating agent with the ability to directly target wt and a set of hotspot mutp53.
General Significance
This work reinforces the encouraging application of SLMP53-1 in the personalized treatment of cancer patients harboring distinct p53 status.

}, keywords = {Cancer, chemotherapy, Mutant, p53, Reactivator}, isbn = {0304-4165}, url = {http://www.sciencedirect.com/science/article/pii/S0304416519302260}, author = {Gomes, Ana Sara and Ramos, Helena and Gomes, Sara and Loureiro, Joana B. and Soares, Joana and Barcherini, Valentina and Monti, Paola and Fronza, Gilberto and Oliveira, Carla and Domingues, Luc{\'\i}lia and Bastos, Margarida and Dourado, Daniel F.A.R. and Ana Lu{\'\i}sa Carvalho and Rom{\~a}o, Maria Jo{\~a}o and Pinheiro, Benedita and Marcelo, Filipa and Alexandra Carvalho and Santos, Maria M.M. and Saraiva, Luc{\'\i}lia} } @article {15380, title = {Towards the mechanistic understanding of enzymatic CO2 reduction}, journal = {ACS CatalysisACS Catalysis}, year = {2020}, note = {

doi: 10.1021/acscatal.0c00086

}, month = {2020}, publisher = {American Chemical Society}, abstract = {

Reducing CO2 is a challenging chemical transformation that biology solves easily, with high efficiency and specificity. In particular, formate dehydrogenases are of great interest since they reduce CO2 to formate, a valuable chemical fuel and hydrogen storage compound. The metal-dependent formate dehydrogenases of prokaryotes can show high activity for CO2 reduction. Here, we report an expression system to produce recombinant W/Sec-FdhAB from Desulfovibrio vulgaris Hildenborough fully loaded with cofactors, its cata-lytic characterization and crystal structures in oxidised and reduced states. The enzyme has very high activi-ty for CO2 reduction and displays remarkable oxygen stability. The crystal structure of the formate-reduced enzyme shows Sec still coordinating the tungsten, supporting a mechanism of stable metal coordination during catalysis. Comparison of the oxidised and reduced structures shows significant changes close to the active site. The DvFdhAB is an excellent model for studying catalytic CO2 reduction and probing the mecha-nism of this conversion.Reducing CO2 is a challenging chemical transformation that biology solves easily, with high efficiency and specificity. In particular, formate dehydrogenases are of great interest since they reduce CO2 to formate, a valuable chemical fuel and hydrogen storage compound. The metal-dependent formate dehydrogenases of prokaryotes can show high activity for CO2 reduction. Here, we report an expression system to produce recombinant W/Sec-FdhAB from Desulfovibrio vulgaris Hildenborough fully loaded with cofactors, its cata-lytic characterization and crystal structures in oxidised and reduced states. The enzyme has very high activi-ty for CO2 reduction and displays remarkable oxygen stability. The crystal structure of the formate-reduced enzyme shows Sec still coordinating the tungsten, supporting a mechanism of stable metal coordination during catalysis. Comparison of the oxidised and reduced structures shows significant changes close to the active site. The DvFdhAB is an excellent model for studying catalytic CO2 reduction and probing the mecha-nism of this conversion.

}, url = {https://doi.org/10.1021/acscatal.0c00086}, author = {Ana Rita Oliveira and Mota, Cristiano and Mourato, Cl{\'a}udia and Domingos, Renato M. and Santos, Marino F. A. and Gesto, Diana and Guigliarelli, Bruno and Santos-Silva, Teresa and Rom{\~a}o, Maria Jo{\~a}o and Pereira, In{\^e}s Antunes Cardoso} } @article {https://doi.org/10.1111/febs.15162, title = {Molecular basis for the preferential recognition of β1,3-1,4-glucans by the family 11 carbohydrate-binding module from Clostridium~thermocellum}, journal = {The FEBS Journal}, volume = {287}, number = {13}, year = {2020}, note = {n/a}, pages = {2723-2743}, abstract = {

Understanding the specific molecular interactions between proteins and β1,3-1,4-mixed-linked d-glucans is fundamental to harvest the full biological and biotechnological potential of these carbohydrates and of proteins that specifically recognize them. The family 11 carbohydrate-binding module from Clostridium~thermocellum (CtCBM11) is known for its binding preference for β1,3-1,4-mixed-linked over β1,4-linked glucans. Despite the growing industrial interest of this protein for the biotransformation of lignocellulosic biomass, the molecular determinants of its ligand specificity are not well defined. In this report, a combined approach of methodologies was used to unravel, at a molecular level, the ligand recognition of CtCBM11. The analysis of the interaction by carbohydrate microarrays and NMR and the crystal structures of CtCBM11 bound to β1,3-1,4-linked glucose oligosaccharides showed that both the chain length and the position of the β1,3-linkage are important for recognition, and identified the tetrasaccharide Glcβ1,4Glcβ1,4Glcβ1,3Glc sequence as a minimum epitope required for binding. The structural data, along with site-directed mutagenesis and ITC studies, demonstrated the specificity of CtCBM11 for the twisted conformation of β1,3-1,4-mixed-linked glucans. This is mediated by a conformation{\textendash}selection mechanism of the ligand in the binding cleft through CH-π stacking and a hydrogen bonding network, which is dependent not only on ligand chain length, but also on the presence of a β1,3-linkage at the reducing end and at specific positions along the β1,4-linked glucan chain. The understanding of the detailed mechanism by which CtCBM11 can distinguish between linear and mixed-linked β-glucans strengthens its exploitation for the design of new biomolecules with improved capabilities and applications in health and agriculture. Database Structural data are available in the Protein Data Bank under the accession codes 6R3M and 6R31.

}, keywords = {3-1, 4-mixed-linked glucans, carbohydrate specificity, carbohydrate-binding module, cellulosome, Clostridium~thermocellum, β1}, doi = {https://doi.org/10.1111/febs.15162}, url = {https://febs.onlinelibrary.wiley.com/doi/abs/10.1111/febs.15162}, author = {Ribeiro, Diana O. and Viegas, Aldino and Pires, Virg{\'\i}nia M R and Medeiros-Silva, Jo{\~a}o and Bule, Pedro and Chai, Wengang and Marcelo, Filipa and Fontes, Carlos M. G. A. and Cabrita, Eurico J. and Palma, Angelina S. and Ana Lu{\'\i}sa Carvalho} } @article {https://doi.org/10.1002/ejic.202000423, title = {Synthesis, Crystal Structure, and DFT Study of Two New Dinuclear Copper(I) Complexes Bearing Ar-BIAN Ligands Functionalized with NO2 Groups}, journal = {European Journal of Inorganic Chemistry}, volume = {2020}, number = {30}, year = {2020}, note = {n/a}, pages = {2900-2911}, abstract = {

{Two new bis(aryl-imino)-acenaphthene, Ar-BIAN (Ar = 2

}, keywords = {Copper(I), Crystal structure, Density Function Calculations, Functionalized Ar-BIAN, synthesis}, doi = {https://doi.org/10.1002/ejic.202000423}, url = {https://chemistry-europe.onlinelibrary.wiley.com/doi/abs/10.1002/ejic.202000423}, author = {Outis, Mani and Rosa, Vitor and C{\'e}sar A. T. Laia and Jo{\~a}o Carlos Lima and Barroso, S{\'o}nia and Ana Lu{\'\i}sa Carvalho and Maria Jos{\'e} Calhorda and Avil{\'e}s, Teresa} } @article {15229, title = {New insights about the monomer and homodimer structures of the human AOX1}, journal = {Physical Chemistry Chemical Physics}, volume = {21}, year = {2019}, note = {n/a}, month = {2019}, pages = {13545 - 13554}, publisher = {The Royal Society of Chemistry}, abstract = {

Human aldehyde oxidase (hAOX1) is a molybdenum dependent enzyme that plays an important role in the metabolism of various compounds either endogenous or xenobiotics. Due to its promiscuity, hAOX1 plays a major role in the pharmacokinetics of many drugs and therefore has gathered a lot of attention from the scientific community and, particularly, from the pharmaceutical industry. In this work, homology modelling, molecular docking and molecular dynamics simulations were used to study the structure of the monomer and dimer of human AOX. The results with the monomer of hAOX1 allowed to shed some light on the role played by thioridazine and two malonate ions that are co-crystalized in the recent X-ray structure of hAOX1. The results show that these molecules endorse several conformational rearrangements in the binding pocket of the enzyme and these changes have an impact in the active site topology as well as in the stability of the substrate (phthalazine). The results show that the presence of both molecules open two gates located at the entrance of the binding pocket, from which results the flooding of the active site. They also endorse several modifications in the shape of the binding pocket (namely the position of Lys893) that, together with the presence of the solvent molecules, favour the release of the substrate to the solvent. Further insights were also obtained with the assembled homodimer of hAOX1. The allosteric inhibitor (THI) binds closely to the region where the dimerization of both monomers occur. These findings suggest that THI can interfere with protein dimerization.

}, isbn = {1463-9076}, url = {http://dx.doi.org/10.1039/C9CP01040H}, author = {Ferreira, P. and Cerqueira, N. M. F. S. A. and Coelho, C. and Fernandes, P. A. and Rom{\~a}o, M.J. and Ramos, M. J.} } @inbook {16063, title = {Aldehyde Oxidases as Enzymes in Phase I Drug Metabolism}, booktitle = {Pharmaceutical Biocatalysis}, year = {2019}, publisher = {Jenny Stanford Publishing}, organization = {Jenny Stanford Publishing}, address = {New York}, author = {Mota, Cristiano and Santos Silva, Teresa and Terao, Mineko and Garattini, Enrico and Rom{\~a}o, Maria Jo{\~a}o and Leimkuehler, Silke}, editor = {Peter Grunwald} } @article {doi:10.1002/2211-5463.12617, title = {Human aldehyde oxidase (hAOX1): structure determination of the Moco-free form of the natural variant G1269R and biophysical studies of single nucleotide polymorphisms}, journal = {FEBS Open Bio}, volume = {9}, number = {5}, year = {2019}, note = {n/a}, pages = {925-934}, abstract = {

Human aldehyde oxidase (hAOX1) is a molybdenum enzyme with high toxicological importance, but its physiological role is still unknown. hAOX1 metabolizes different classes of xenobiotics and is one of the main drug-metabolizing enzymes in the liver, along with cytochrome P450. hAOX1 oxidizes and inactivates a large number of drug molecules and has been responsible for the failure of several phase I clinical trials. The interindividual variability of drug-metabolizing enzymes caused by single nucleotide polymorphisms (SNPs) is highly relevant in pharmaceutical treatments. In this study, we present the crystal structure of the inactive variant G1269R, revealing the first structure of a molybdenum cofactor (Moco)-free form of hAOX1. These data allowed to model, for the first time, the flexible Gate 1 that controls access to the active site. Furthermore, we inspected the thermostability of wild-type hAOX1 and hAOX1 with various SNPs (L438V, R1231H, G1269R or S1271L) by CD spectroscopy and ThermoFAD, revealing that amino acid exchanges close to the Moco site can impact protein stability up to 10~{\textdegree}C. These results correlated with biochemical and structural data and enhance our understanding of hAOX1 and the effect of SNPs in the gene encoding this enzyme in the human population. Enzymes Aldehyde oxidase (EC1.2.3.1); xanthine dehydrogenase (EC1.17.1.4); xanthine oxidase (EC1.1.3.2). Databases Structural data are available in the Protein Data Bank under the accession number 6Q6Q.

}, keywords = {human aldehyde oxidase, molybdenum cofactor, single nucleotide polymorphism, Xanthine Oxidase}, doi = {10.1002/2211-5463.12617}, url = {https://febs.onlinelibrary.wiley.com/doi/abs/10.1002/2211-5463.12617}, author = {Mota, Cristiano and Esmaeeli, Mariam and Coelho, Catarina and Santos-Silva, Teresa and Wolff, Martin and Foti, Alessandro and Leimk{\"u}hler, Silke and Rom{\~a}o, Maria Jo{\~a}o} } @article {doi:10.1002/prp2.538, title = {Systematic exploration of predicted destabilizing nonsynonymous single nucleotide polymorphisms (nsSNPs) of human aldehyde oxidase: A Bio-informatics study}, journal = {Pharmacology Research \& Perspectives}, volume = {7}, number = {6}, year = {2019}, note = {n/a}, pages = {e00538}, abstract = {

Abstract Aldehyde Oxidase (hAOX1) is a cytosolic enzyme involved in the metabolism of drugs and xenobiotic compounds. The enzyme belongs to the xanthine oxidase (XO) family of Mo containing enzyme and is a homo-dimer of two 150~kDa monomers. Nonsynonymous Single Nucleotide Polymorphisms (nsSNPs) of hAOX1 have been reported as affecting the ability of the enzyme to metabolize different substrates. Some of these nsSNPs have been biochemically and structurally characterized but the lack of a systematic and comprehensive study regarding all described and validated nsSNPs is urgent, due to the increasing importance of the enzyme in drug development, personalized medicine and therapy, as well as in pharmacogenetic studies. The objective of the present work was to collect all described nsSNPs of hAOX1 and utilize a series of bioinformatics tools to predict their effect on protein structure stability with putative implications on phenotypic functional consequences. Of 526 nsSNPs reported in NCBI-dbSNP, 119 are identified as deleterious whereas 92 are identified as nondeleterious variants. The stability analysis was performed for 119 deleterious variants and the results suggest that 104 nsSNPs may be responsible for destabilizing the protein structure, whereas five variants may increase the protein stability. Four nsSNPs do not have any impact on protein structure (neutral nsSNPs) of hAOX1. The prediction results of the remaining six nsSNPs are nonconclusive. The in silico results were compared with available experimental data. This methodology can also be used to identify and prioritize the stabilizing and destabilizing variants in other enzymes involved in drug metabolism.

}, keywords = {Aldehyde Oxidase, computational genomic, in silico analysis, pharmacogenetic, single nucleotide polymorphism}, doi = {10.1002/prp2.538}, url = {https://bpspubs.onlinelibrary.wiley.com/doi/abs/10.1002/prp2.538}, author = {Coelho, Catarina and Muthukumaran, Jayaraman and Santos-Silva, Teresa and Rom{\~a}o, Maria Jo{\~a}o} } @article {Carvalho2018, title = {{CHAPTER 2 Structural Elucidation of Macromolecules}}, journal = {Essential Techniques for Medical and Life Scientists: A Guide to Contemporary Methods and Current Applications with the Protocols}, year = {2018}, note = {n/a}, month = {sep}, pages = {30{\textendash}91}, publisher = {BENTHAM SCIENCE PUBLISHERS}, abstract = {n/a}, keywords = {cryo-em, Drug Design, ligand interactions, macromolecular structure, NMR, protein-, saxs, X-ray crystallography}, doi = {10.2174/9781681087092118010005}, url = {http://www.eurekaselect.com/node/165742}, author = {Ana Lu{\'\i}sa Carvalho and Santos-Silva, Teresa and Rom{\~a}o, Maria Jo{\~a}o and Eurico, J and Marcelo, Filipa} } @article {13723, title = {Critical overview on the structure and metabolism of human aldehyde oxidase and its role in pharmacokinetics}, volume = {368}, year = {2018}, note = {n/a}, month = {2018}, pages = {35 - 59}, abstract = {

Aldehyde oxidases are molybdenum and flavin dependent enzymes characterized by a very wide substrate specificity and performing diverse reactions that include oxidations (e.g., aldehydes and aza-heterocycles), hydrolysis of amide bonds, and reductions (e.g., nitro, S-oxides and N-oxides). Oxidation reactions and amide hydrolysis occur at the molybdenum site while the reductions are proposed to occur at the flavin site. AOX activity affects the metabolism of different drugs and xenobiotics, some of which designed to resist other liver metabolizing enzymes (e.g., cytochrome P450 monooxygenase isoenzymes), raising its importance in drug development. This work consists of a comprehensive overview on aldehyde oxidases, concerning the genetic evolution of AOX, its diversity among the human population, the crystal structures available, the known catalytic reactions and the consequences in pre-clinical pharmacokinetic and pharmacodynamic studies. Analysis of the different animal models generally used for pre-clinical trials and comparison between the human (hAOX1), mouse homologs as well as the related xanthine oxidase (XOR) are extensively considered. The data reviewed also include a systematic analysis of representative classes of molecules that are hAOX1 substrates as well as of typical and well characterized hAOX1 inhibitors. The considerations made on the basis of a structural and functional analysis are correlated with reported kinetic and metabolic data for typical classes of drugs, searching for potential structural determinants that may dictate substrate and/or inhibitor specificities.

}, keywords = {Aldehyde Oxidase, Drug metabolism, Hepatic clearance, Molybdoenzymes, Non-CYP enzymes, Xenobiotics}, isbn = {0010-8545}, url = {http://www.sciencedirect.com/science/article/pii/S0010854517306185}, author = {Mota, Cristiano and Coelho, Catarina and Leimk{\"u}hler, Silke and Garattini, Enrico and Terao, Mineko and Santos-Silva, Teresa and Rom{\~a}o, Maria Jo{\~a}o} } @article {13166, title = {First insights of peptidoglycan amidation in Gram-positive bacteria - the high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD}, journal = {Scientific Reports}, volume = {8}, year = {2018}, note = {

n/a

}, month = {2018}, pages = {5313}, abstract = {

Gram-positive bacteria homeostasis and antibiotic resistance mechanisms are dependent on the intricate architecture of the cell wall, where amidated peptidoglycan plays an important role. The amidation reaction is carried out by the bi-enzymatic complex MurT-GatD, for which biochemical and structural information is very scarce. In this work, we report the first crystal structure of the glutamine amidotransferase member of this complex, GatD from Staphylococcus aureus, at 1.85 {\r A} resolution. A glutamine molecule is found close to the active site funnel, hydrogen-bonded to the conserved R128. In vitro functional studies using 1H-NMR spectroscopy showed that S. aureus MurT-GatD complex has glutaminase activity even in the absence of lipid II, the MurT substrate. In addition, we produced R128A, C94A and H189A mutants, which were totally inactive for glutamine deamidation, revealing their essential role in substrate sequestration and catalytic reaction. GatD from S. aureus and other pathogenic bacteria share high identity to enzymes involved in cobalamin biosynthesis, which can be grouped in a new sub-family of glutamine amidotransferases. Given the ubiquitous presence of GatD, these results provide significant insights into the molecular basis of the so far undisclosed amidation mechanism, contributing to the development of alternative therapeutics to fight infections.

}, isbn = {2045-2322}, url = {https://doi.org/10.1038/s41598-018-22986-3}, author = {Leisico, Francisco and V. Vieira, Diana and Figueiredo, Teresa A. and Silva, Micael and Cabrita, Eurico J. and Sobral, Rita G. and Ludovice, Ana Madalena and Trinc{\~a}o, Jos{\'e} and Rom{\~a}o, Maria Jo{\~a}o and de Lencastre, Herm{\'\i}nia and Santos-Silva, Teresa} } @article {EJOC:EJOC201701500, title = {Synthesis, cytotoxicity evaluation in human cell lines and in vitro DNA interaction of a hetero arylidene-9(10H)-anthrone}, journal = {European Journal of Organic Chemistry}, year = {2018}, note = {

n/a

}, pages = {n/a{\textendash}n/a}, abstract = {

A new and never yet reported hetero arylidene-9(10H)-anthrone structure (4) was unexpectedly isolated on reaction of 1,2-dimethyl-3-ethylimidazolium iodide (2) and 9-anthracenecarboxaldehyde (3) under basic conditions. Its structure was unequivocally attributed by X-ray crystallography. No cytotoxicity in human healthy fibroblasts and in two different cancer cell lines was observed indicating its applicability in biological systems. Compound 4 interacts with CT-DNA by intercalation between the adjacent base pairs of DNA with a high binding affinity (Kb = 2.0({\textpm} 0.20) x 105 M-1) which is 10x higher than that described for doxorubicin (Kb = 3.2 ({\textpm}0.23) {\texttimes} 104 M-1). Furthermore, compound 4 quenches the fluorescence emission of GelRed-CT-DNA system with a quenching constant (KSV) of 3.3({\textpm}0.3) x 103 M-1 calculated by the Stern-Volmer equation.

}, keywords = {anthracenecarboxaldehyde, arylidene anthrone, Decarbonylation, DNA intercalation, Imidazolium salt}, issn = {1099-0690}, doi = {10.1002/ejoc.201701500}, url = {http://dx.doi.org/10.1002/ejoc.201701500}, author = {Branco, Paula S. and Peixoto, Daniela and Figueiredo, Margarida and Malta, Gabriela and Catarina Roma-Rodrigues and Batista, Pedro Viana and Alexandra R. Fernandes and Barroso, S{\'o}nia and Carvalho, Ana Luisa and Afonso, Carlos A. M. and Ferreira, Lu{\'\i}sa Maria} } @article {ijms19041184, title = {The Crystal Structure of the R280K Mutant of Human p53 Explains the Loss of DNA Binding}, journal = {International Journal of Molecular Sciences}, volume = {19}, number = {4}, ARTICLE NUMBER = {1184}, year = {2018}, note = {n/a}, abstract = {

The p53 tumor suppressor is widely found to be mutated in human cancer. This protein is regarded as a molecular hub regulating different cell responses, namely cell death. Compelling data have demonstrated that the impairment of p53 activity correlates with tumor development and maintenance. For these reasons, the reactivation of p53 function is regarded as a promising strategy to halt cancer. In the present work, the recombinant mutant p53R280K DNA binding domain (DBD) was produced for the first time, and its crystal structure was determined in the absence of DNA to a resolution of 2.0 {\r A}. The solved structure contains four molecules in the asymmetric unit, four zinc(II) ions, and 336 water molecules. The structure was compared with the wild-type p53 DBD structure, isolated and in complex with DNA. These comparisons contributed to a deeper understanding of the mutant p53R280K structure, as well as the loss of DNA binding related to halted transcriptional activity. The structural information derived may also contribute to the rational design of mutant p53 reactivating molecules with potential application in cancer treatment.

}, issn = {1422-0067}, doi = {10.3390/ijms19041184}, url = {http://www.mdpi.com/1422-0067/19/4/1184}, author = {Gomes, Ana Sara and Trov{\~a}o, Filipa and Andrade Pinheiro, Benedita and Freire, Filipe and Gomes, Sara and Oliveira, Carla and Domingues, Luc{\'\i}lia and Rom{\~a}o, Maria Jo{\~a}o and Saraiva, Luc{\'\i}lia and Ana Lu{\'\i}sa Carvalho} } @article {Kumar2018, title = {Novel insights into the degradation of β-1,3-glucans by the cellulosome of Clostridium thermocellum revealed by structure and function studies of a family 81 glycoside hydrolase}, journal = {International Journal of Biological Macromolecules}, year = {2018}, note = {n/a}, pages = {-}, abstract = {

Abstract The family 81 glycoside hydrolase (GH81) from Clostridium thermocellum is a β-1,3-glucanase belonging to cellulosomal complex. The gene encoding \{GH81\} from Clostridium thermocellum (CtLam81A) was cloned and expressed displaying a molecular mass of ~82 kDa. CtLam81A showed maximum activity against laminarin (100 U/mg), followed by curdlan (65 U/mg), at pH 7.0 and 75 {\textdegree}C. CtLam81A displayed Km, 2.1 {\textpm} 0.12 mg/ml and Vmax, 109 {\textpm} 1.8 U/mg, against laminarin under optimized conditions. CtLam81A activity was significantly enhanced by Ca2+ or Mg2+ ions. Melting curve analysis of CtLam81A showed an increase in melting temperature from 91 {\textdegree}C to 96 {\textdegree}C by Ca2+ or Mg2+ ions and decreased to 82 {\textdegree}C by EDTA, indicating that Ca2+ and Mg2+ ions may be involved in catalysis and in maintaining structural integrity. \{TLC\} and MALDI-TOF analysis of β-1,3-glucan hydrolysed products released initially, showed β-1,3-glucan-oligosaccharides degree of polymerization (DP) from \{DP2\} to DP7, confirming an endo-mode of action. The catalytically inactive mutant CtLam81A-E515A generated by site-directed mutagenesis was co-crystallized and tetragonal crystals diffracting up to 1.4 {\r A} resolution were obtained. CtLam81A-E515A contained 15 α-helices and 38 β-strands forming a four-domain structure viz. a β-sandwich domain I at N-terminal, an α/β-domain II, an (α/α)6 barrel domain III, and a small 5-stranded β-sandwich domain IV.

}, keywords = {X-ray crystallography}, issn = {0141-8130}, doi = {https://doi.org/10.1016/j.ijbiomac.2018.06.003}, url = {https://www.sciencedirect.com/science/article/pii/S0141813018322384}, author = {Krishan Kumar and Correia, M{\'a}rcia and Virg{\'\i}nia R. Pires and Arun Dhillon and Kedar Sharma and Vikky Rajulapati and Fontes, Carlos M. G. A. and Ana Lu{\'\i}sa Carvalho and Goyal, Arun} } @inbook {10.1039/9781788010641-00159, title = {Targeting protein-carbohydrate interactions in plant cell-wall biodegradation: the power of carbohydrate microarrays}, booktitle = {Carbohydrate Chemistry: Chemical and Biological Approaches Volume 43}, volume = {43}, year = {2018}, note = {n/a}, pages = {159-176}, publisher = {The Royal Society of Chemistry}, organization = {The Royal Society of Chemistry}, abstract = {

The plant cell-wall is constituted by structurally diverse polysaccharides. The biodegradation of these is a crucial process for life sustainability. Cellulolytic microorganisms are highly efficient in this process by assembling modular architectures of carbohydrate-active enzymes with appended non-catalytic carbohydrate-binding modules (CBMs). Carbohydrate microarrays offer high-throughput and sensitive tools for uncovering carbohydrate-binding specificities of CBMs{,} which is pivotal to understand the function of these modules in polysaccharide biodegradation mechanisms. Features of this technology will be here briefly reviewed with highlights of microarray approaches to study plant-carbohydrates and CBM-carbohydrate interactions{,} along with an overview of plant polysaccharides and microorganisms strategies for their recognition.

}, isbn = {978-1-78801-003-0}, doi = {10.1039/9781788010641-00159}, url = {http://dx.doi.org/10.1039/9781788010641-00159}, author = {Ribeiro, Diana O. and Pinheiro, Benedita A. and Carvalho, Ana Luisa and Palma, Angelina S.} } @article {Kryshtafovych2017, title = {Target highlights from the first post-PSI CASP experiment (CASP12, May-August 2016)}, journal = {Proteins: Structure, Function, and Bioinformatics}, year = {2017}, note = {

n/a

}, month = {oct}, abstract = {

The functional and biological significance of the selected CASP12 targets are described by the authors of the structures. The crystallographers discuss the most interesting structural features of the target proteins and assess whether these features were correctly reproduced in the predictions submitted to the CASP12 experiment. This article is protected by copyright. All rights reserved.

}, keywords = {CASP, NMR, protein structure prediction, X-ray crystallography}, issn = {08873585}, doi = {10.1002/prot.25392}, url = {http://onlinelibrary.wiley.com/doi/10.1002/prot.25392/abstract}, author = {Kryshtafovych, Andriy and Albrecht, Reinhard and Basl{\'e}, Arnaud and Bule, Pedro and Caputo, Alessandro T. and Carvalho, Ana Luisa and Chao, Kinlin L. and Diskin, Ron and Fidelis, Krzysztof and Fontes, Carlos M. G. A. and Fredslund, Folmer and Gilbert, Harry J. and Goulding, Celia W. and Hartmann, Marcus D. and Hayes, Christopher S. and Herzberg, Osnat and Hill, Johan C. and Joachimiak, Andrzej and Kohring, Gert-Wieland and Koning, Roman I. and {Lo Leggio}, Leila and Mangiagalli, Marco and Michalska, Karolina and Moult, John and Najmudin, Shabir and Nardini, Marco and Nardone, Valentina and Ndeh, Didier and Nguyen, Thanh H. and Pintacuda, Guido and Postel, Sandra and van Raaij, Mark J. and Roversi, Pietro and Shimon, Amir and Singh, Abhimanyu K. and Sundberg, Eric J. and Tars, Kaspars and Zitzmann, Nicole and Schwede, Torsten} } @article {Pires2017a, title = {Stability and ligand promiscuity of type A carbohydrate-binding modules are illustrated by the structure of Spirochaeta thermophila StCBM64C}, journal = {Journal of Biological Chemistry}, volume = {292}, number = {12}, year = {2017}, note = {

n/a

}, month = {mar}, pages = {4847{\textendash}4860}, abstract = {

Deconstruction of cellulose, the most abundant plant cell wall polysaccharide, requires the cooperative activity of a large repertoire of microbial enzymes. Modular cellulases contain non-catalytic type A Carbohydrate-Binding Modules (CBMs) that specifically bind to the crystalline regions of cellulose, thus promoting enzyme efficacy through proximity and targeting effects. Although type A CBMs play a critical role in cellulose recycling, their mechanism of action remains poorly understood. Here we produced a library of recombinant CBMs representative of the known diversity of type A modules. The binding properties of 40 CBMs, in fusion with an N-terminal green fluorescence protein (GFP) domain, revealed that type A CBMs possess the ability to recognize different crystalline forms of cellulose and chitin over a wide range of temperatures, pHs and ionic strengths. A Spirochaeta thermophila CBM64, in particular, displayed plasticity in its capacity to bind both crystalline and soluble carbohydrates under a wide range of extreme conditions. The structure of S. thermophila StCBM64C revealed an untwisted, flat, carbohydrate-binding interface comprising the side chains of four tryptophan residues in a coplanar linear arrangement. Significantly, two highly conserved asparagine side chains, each one located between two tryptophan residues, are critical to insoluble and soluble glucan recognition but not to bind xyloglucan. Thus, CBM64 compact structure and its extended and versatile ligand interacting platform illustrates how type A CBMs target their appended plant cell wall degrading enzymes to a diversity of recalcitrant carbohydrates under a wide range of environmental conditions.

}, issn = {1083351X}, doi = {10.1074/jbc.M116.767541}, url = {http://europepmc.org/abstract/med/28179427}, author = {Pires, Virg{\'\i}nia M R and Pereira, Pedro M M and Br{\'a}s, Joana L A and Correia, M{\'a}rcia and Cardoso, V{\^a}nia and Bule, Pedro and Alves, Victor D. and Najmudin, Shabir and Venditto, Immacolata and Ferreira, Lu{\'\i}s M. A. and Rom{\~a}o, Maria Jo{\~a}o and Ana Lu{\'\i}sa Carvalho and Fontes, Carlos M. G. A. and Prazeres, Duarte Miguel} } @article {11734, title = {Renaissance of protein crystallization and precipitation in biopharmaceuticals purification}, journal = {Biotechnology Advances}, volume = {35}, year = {2017}, note = {

n/a

}, month = {2017/1//}, pages = {41 - 50}, abstract = {

AbstractThe current chromatographic approaches used in protein purification are not keeping pace with the increasing biopharmaceutical market demand. With the upstream improvements, the bottleneck shifted towards the downstream process. New approaches rely in Anything But Chromatography methodologies and revisiting former techniques with a bioprocess perspective. Protein crystallization and precipitation methods are already implemented in the downstream process of diverse therapeutic biological macromolecules, overcoming the current chromatographic bottlenecks. Promising work is being developed in order to implement crystallization and precipitation in the purification pipeline of high value therapeutic molecules. This review focuses in the role of these two methodologies in current industrial purification processes, and highlights their potential implementation in the purification pipeline of high value therapeutic molecules, overcoming chromatographic holdups.

}, keywords = {Anything but chromatography, Biopharmaceutical, Crystallization, Downstream process, precipitation, Protein Purification}, isbn = {0734-9750}, url = {http://www.sciencedirect.com/science/article/pii/S0734975016301513}, author = {Raquel dos Santos and Ana Lu{\'\i}sa Carvalho and Roque, A. Cec{\'\i}lia A.} } @article {11936, title = {Highly selective tungstate transporter protein TupA from Desulfovibrio alaskensis G20}, journal = {Scientific Reports}, volume = {7}, year = {2017}, note = {

n/a

}, month = {2017}, pages = {5798}, abstract = {

Molybdenum and tungsten are taken up by bacteria and archaea as their soluble oxyanions through high affinity transport systems belonging to the ATP-binding cassette (ABC) transporters. The component A (ModA/TupA) of these transporters is the first selection gate from which the cell differentiates between MoO4 2-, WO4 2- and other similar oxyanions. We report the biochemical characterization and the crystal structure of the apo-TupA from Desulfovibrio desulfuricans G20, at 1.4 {\r A} resolution. Small Angle X-ray Scattering data suggests that the protein adopts a closed and more stable conformation upon ion binding. The role of the arginine 118 in the selectivity of the oxyanion was also investigated and three mutants were constructed: R118K, R118E and R118Q. Isothermal titration calorimetry clearly shows the relevance of this residue for metal discrimination and oxyanion binding. In this sense, the three variants lost the ability to coordinate molybdate and the R118K mutant keeps an extremely high affinity for tungstate. These results contribute to an understanding of the metal-protein interaction, making it a suitable candidate for a recognition element of a biosensor for tungsten detection.

}, isbn = {2045-2322}, url = {https://doi.org/10.1038/s41598-017-06133-y}, author = {Otrelo-Cardoso, Ana Rita and Nair, Rashmi R and Correia, M{\'a}rcia A. S. and Cordeiro, Raquel S. Correia and Panjkovich, Alejandro and Svergun, Dmitri I. and Santos-Silva, Teresa and Rivas, Maria G.} } @article {11942, title = {Ion-Exchange Membranes for Stable Derivatization of Protein Crystals}, journal = {Crystal Growth \& DesignCrystal Growth \& Design}, year = {2017}, note = {

doi: 10.1021/acs.cgd.7b00315

}, month = {2017}, publisher = {American Chemical Society}, abstract = {n/a}, isbn = {1528-7483}, url = {http://dx.doi.org/10.1021/acs.cgd.7b00315}, author = {Polino, Mariella and Carvalho, Ana Lu{\i}́sa and Juknaite, Lina and Portugal, Carla A. M. and Coelhoso, Isabel M. and Rom{\~a}o, Maria Jo{\~a}o and Jo{\~a}o G. Crespo} } @article {WATSON2017865, title = {Electron transfer through arsenite oxidase: Insights into Rieske interaction with cytochrome c}, journal = {Biochimica et Biophysica Acta (BBA) - Bioenergetics}, volume = {1858}, number = {10}, year = {2017}, note = {n/a}, pages = {865 - 872}, abstract = {

Arsenic is a widely distributed environmental toxin whose presence in drinking water poses a threat to \>140 million people worldwide. The respiratory enzyme arsenite oxidase from various bacteria catalyses the oxidation of arsenite to arsenate and is being developed as a biosensor for arsenite. The arsenite oxidase from Rhizobium sp. str. NT-26 (a member of the Alphaproteobacteria) is a heterotetramer consisting of a large catalytic subunit (AioA), which contains a molybdenum centre and a 3Fe-4S cluster, and a small subunit (AioB) containing a Rieske 2Fe-2S cluster. Stopped-flow spectroscopy and isothermal titration calorimetry (ITC) have been used to better understand electron transfer through the redox-active centres of the enzyme, which is essential for biosensor development. Results show that oxidation of arsenite at the active site is extremely fast with a rate of \>4000s-1 and reduction of the electron acceptor is rate-limiting. An AioB-F108A mutation results in increased activity with the artificial electron acceptor DCPIP and decreased activity with cytochrome c, which in the latter as demonstrated by ITC is not due to an effect on the protein-protein interaction but instead to an effect on electron transfer. These results provide further support that the AioB F108 is important in electron transfer between the Rieske subunit and cytochrome c and its absence in the arsenite oxidases from the Betaproteobacteria may explain the inability of these enzymes to use this electron acceptor.

}, keywords = {Arsenite oxidase, Cytochrome, Isothermal titration calorimetry, Molybdenum enzyme, Rate-limiting step, Rieske protein, Stopped-flow spectroscopy}, issn = {0005-2728}, doi = {https://doi.org/10.1016/j.bbabio.2017.08.003}, url = {http://www.sciencedirect.com/science/article/pii/S0005272817301160}, author = {Cameron Watson and Dimitri Niks and Russ Hille and Marta Vieira and Barbara Schoepp-Cothenet and Alexandra T. Marques and Rom{\~a}o, Maria Jo{\~a}o and Santos-Silva, Teresa and Joanne M. Santini} } @article {Kowacz2017362, title = {Infrared light-induced protein crystallization. Structuring of protein interfacial water and periodic self-assembly}, journal = {Journal of Crystal Growth}, volume = {457}, year = {2017}, note = {

SI: CRYS_ECCG5

}, pages = {362 - 368}, abstract = {

Abstract We show that a physical trigger, a non-ionizing infrared (IR) radiation at wavelengths strongly absorbed by liquid water, can be used to induce and kinetically control protein (periodic) self-assembly in solution. This phenomenon is explained by considering the effect of {\.I}R\} light on the structuring of protein interfacial water. Our results indicate that the {\.I}R\} radiation can promote enhanced mutual correlations of water molecules in the protein hydration shell. We report on the radiation-induced increase in both the strength and cooperativeness of H-bonds. The presence of a structured dipolar hydration layer can lead to attractive interactions between like-charged biomacromolecules in solution (and crystal nucleation events). Furthermore, our study suggests that enveloping the protein within a layer of structured solvent (an effect enhanced by {\.I}R\} light) can prevent the protein non-specific aggregation favoring periodic self-assembly. Recognizing the ability to affect protein-water interactions by means of {\.I}R\} radiation may have important implications for biological and bio-inspired systems.

}, keywords = {A4. Crystal growth from solution}, issn = {0022-0248}, doi = {https://doi.org/10.1016/j.jcrysgro.2016.01.003}, url = {http://www.sciencedirect.com/science/article/pii/S0022024816000178}, author = {Kowacz, Magdalena and Mateusz Marchel and Lina Juknait{\'e} and Jos{\'e} M.S.S. Esperan{\c c}a and Rom{\~a}o, Maria Jo{\~a}o and Ana Lu{\'\i}sa Carvalho and Lu{\'\i}s Paulo N. Rebelo} } @article {Rom{\~a}o201739, title = {Structural basis for the role of mammalian aldehyde oxidases in the metabolism of drugs and xenobiotics}, journal = {Current Opinion in Chemical Biology}, volume = {37}, year = {2017}, note = {

Biocatalysis \& biotransformation * Bioinorganic Chemistry

}, pages = {39 - 47}, abstract = {

Aldehyde oxidases (AOXs) are molybdo-flavoenzymes characterized by broad substrate specificity, oxidizing aromatic/aliphatic aldehydes into the corresponding carboxylic acids and hydroxylating various heteroaromatic rings. Mammals are characterized by a complement of species-specific \{AOX\} isoenzymes, that varies from one in humans (AOX1) to four in rodents (AOX1, AOX2, \{AOX3\} and AOX4). The physiological function of mammalian \{AOX\} isoenzymes is unknown, although human \{AOX1\} is an emerging enzyme in phase-I drug metabolism. Indeed, the number of therapeutic molecules under development which act as \{AOX\} substrates is increasing. The recent crystallization and structure determination of human \{AOX1\} as well as mouse \{AOX3\} has brought new insights into the mechanisms underlying substrate/inhibitor binding as well as the catalytic activity of this class of enzymes.

}, issn = {1367-5931}, doi = {https://doi.org/10.1016/j.cbpa.2017.01.005}, url = {http://www.sciencedirect.com/science/article/pii/S1367593117300066}, author = {Rom{\~a}o, Maria Jo{\~a}o and Coelho, Catarina and Santos-Silva, Teresa and Foti, Alessandro and Terao, Mineko and Garattini, Enrico and Leimk{\"u}hler, Silke} } @article {ADFM:ADFM201700803, title = {Tunable Gas Sensing Gels by Cooperative Assembly}, journal = {Advanced Functional Materials}, volume = {27}, number = {27}, year = {2017}, note = {

1700803

}, pages = {1700803{\textendash}n/a}, abstract = {

The cooperative assembly of biopolymers and small molecules can yield functional materials with precisely tunable properties. Here, the fabrication, characterization, and use of multicomponent hybrid gels as selective gas sensors are reported. The gels are composed of liquid crystal droplets self-assembled in the presence of ionic liquids, which further coassemble with biopolymers to form stable matrices. Each individual component can be varied and acts cooperatively to tune gels{\textquoteright} structure and function. The unique molecular environment in hybrid gels is explored for supramolecular recognition of volatile compounds. Gels with distinct compositions are used as optical and electrical gas sensors, yielding a combinatorial response conceptually mimicking olfactory biological systems, and tested to distinguish volatile organic compounds and to quantify ethanol in automotive fuel. The gel response is rapid, reversible, and reproducible. These robust, versatile, modular, pliant electro-optical soft materials possess new possibilities in sensing triggered by chemical and physical stimuli.

}, keywords = {gas sensing, gelatin, ionic liquids, liquid crystals, self-assembly}, issn = {1616-3028}, doi = {10.1002/adfm.201700803}, url = {http://dx.doi.org/10.1002/adfm.201700803}, author = {Hussain, Abid and Semeano, Ana T. S. and Susana I C J Palma and Ana S Pina and Almeida, Jos{\'e} and Medrado, B{\'a}rbara F. and P{\'a}dua, Ana C. C. S. and Carvalho, Ana L. and Madalena Dion{\'\i}sio and Li, Rosamaria W. C. and Gamboa, Hugo and Ulijn, Rein V. and Gruber, Jonas and Roque, Ana C A} } @article {Correia2016, title = {{The Escherichia coli Periplasmic Aldehyde Oxidoreductase Is an Exceptional Member of the Xanthine Oxidase Family of Molybdoenzymes}}, journal = {ACS Chemical Biology}, volume = {11}, number = {10}, year = {2016}, note = {

n/a

}, month = {oct}, pages = {2923{\textendash}2935}, abstract = {

The xanthine oxidase (XO) family comprises molybdenum-dependent enzymes that usually form homodimers (or dimers of heterodimers/trimers) organized in three domains that harbor two [2Fe-2S] clusters, one FAD, and a Mo cofactor. In this work, we crystallized an unusual member of the family, the periplasmic aldehyde oxidoreductase PaoABC from Escherichia coli. This is the first example of an E. coli protein containing a molybdopterin-cytosine-dinucleotide cofactor and is the only heterotrimer of the XO family so far structurally characterized. The crystal structure revealed the presence of an unexpected [4Fe-4S] cluster, anchored to an additional 40 residues subdomain. According to phylogenetic analysis, proteins containing this cluster are widely spread in many bacteria phyla, putatively through repeated gene transfer events. The active site of PaoABC is highly exposed to the surface with no aromatic residues and an arginine (PaoC-R440) making a direct interaction with PaoC-E692, which acts as a base catalyst. In order to understand the importance of R440, kinetic assays were carried out, and the crystal structure of the PaoC-R440H variant was also determined.

}, issn = {1554-8929}, doi = {10.1021/acschembio.6b00572}, url = {http://www.ncbi.nlm.nih.gov/pubmed/27622978 http://pubs.acs.org/doi/abs/10.1021/acschembio.6b00572}, author = {Correia, M{\'a}rcia A. S. and Otrelo-Cardoso, Ana Rita and Schwuchow, Viola and {Sigfridsson Clauss}, Kajsa G. V. and Haumann, Michael and Rom{\~a}o, Maria Jo{\~a}o and Leimk{\"u}hler, Silke and Santos-Silva, Teresa} } @article {Bras2016, title = {Diverse specificity of cellulosome attachment to the bacterial cell surface}, journal = {Scientific Reports}, volume = {6}, year = {2016}, note = {

n/a

}, month = {dec}, pages = {38292}, publisher = {The Author(s)}, abstract = {

During the course of evolution, the cellulosome, one of Nature{\textquoteright}s most intricate multi-enzyme complexes, has been continuously fine-tuned to efficiently deconstruct recalcitrant carbohydrates. To facilitate the uptake of released sugars, anaerobic bacteria use highly ordered protein-protein interactions to recruit these nanomachines to the cell surface. Dockerin modules located within a non-catalytic macromolecular scaffold, whose primary role is to assemble cellulosomal enzymatic subunits, bind cohesin modules of cell envelope proteins, thereby anchoring the cellulosome onto the bacterial cell. Here we have elucidated the unique molecular mechanisms used by anaerobic bacteria for cellulosome cellular attachment. The structure and biochemical analysis of five cohesin-dockerin complexes revealed that cell surface dockerins contain two cohesin-binding interfaces, which can present different or identical specificities. In contrast to the current static model, we propose that dockerins utilize multivalent modes of cohesin recognition to recruit cellulosomes to the cell surface, a mechanism that maximises substrate access while facilitating complex assembly.

}, url = {http://dx.doi.org/10.1038/srep38292 http://10.1038/srep38292 http://www.nature.com/articles/srep38292{\#}supplementary-information}, author = {Br{\'a}s, Joana L A and Pinheiro, Benedita A. and Cameron, Kate and Cuskin, Fiona and Viegas, Aldino and Najmudin, Shabir and Bule, Pedro and Pires, Virginia M R and Rom{\~a}o, Maria Jo{\~a}o and Bayer, Edward A. and Spencer, Holly L and Smith, Steven and Gilbert, Harry J. and Alves, Victor D. and Ana Lu{\'\i}sa Carvalho and Fontes, Carlos M. G. A.} } @article {Foti1277, title = {Optimization of the Expression of Human Aldehyde Oxidase for Investigations of Single-Nucleotide Polymorphisms}, journal = {Drug Metabolism and Disposition}, volume = {44}, number = {8}, year = {2016}, note = {n/a}, pages = {1277{\textendash}1285}, publisher = {American Society for Pharmacology and Experimental Therapeutics}, abstract = {

Aldehyde oxidase (AOX1) is an enzyme with broad substrate specificity, catalyzing the oxidation of a wide range of endogenous and exogenous aldehydes as well as N-heterocyclic aromatic compounds. In humans, the enzyme{\textquoteright}s role in phase I drug metabolism has been established and its importance is now emerging. However, the true physiologic function of AOX1 in mammals is still unknown. Further, numerous single-nucleotide polymorphisms (SNPs) have been identified in human AOX1. SNPs are a major source of interindividual variability in the human population, and SNP-based amino acid exchanges in AOX1 reportedly modulate the catalytic function of the enzyme in either a positive or negative fashion. For the reliable analysis of the effect of amino acid exchanges in human proteins, the existence of reproducible expression systems for the production of active protein in ample amounts for kinetic, spectroscopic, and crystallographic studies is required. In our study we report an optimized expression system for hAOX1 in Escherichia coli using a codon-optimized construct. The codon-optimization resulted in an up to 15-fold increase of protein production and a simplified purification procedure. The optimized expression system was used to study three SNPs that result in amino acid changes C44W, G1269R, and S1271L. In addition, the crystal structure of the S1271L SNP was solved. We demonstrate that the recombinant enzyme can be used for future studies to exploit the role of AOX in drug metabolism, and for the identification and synthesis of new drugs targeting AOX when combined with crystallographic and modeling studies.

}, issn = {0090-9556}, doi = {10.1124/dmd.115.068395}, url = {http://dmd.aspetjournals.org/content/44/8/1277}, author = {Foti, Alessandro and Hartmann, Tobias and Coelho, Catarina and Santos-Silva, Teresa and Rom{\~a}o, Maria Jo{\~a}o and Leimk{\"u}hler, Silke} } @article {Terao2016, title = {Structure and function of mammalian aldehyde oxidases}, journal = {Archives of Toxicology}, volume = {90}, number = {4}, year = {2016}, note = {n/a}, pages = {753{\textendash}780}, abstract = {

Mammalian aldehyde oxidases (AOXs; EC1.2.3.1) are a group of conserved proteins belonging to the family of molybdo-flavoenzymes along with the structurally related xanthine dehydrogenase enzyme. AOXs are characterized by broad substrate specificity, oxidizing not only aromatic and aliphatic aldehydes into the corresponding carboxylic acids, but also hydroxylating a series of heteroaromatic rings. The number of AOX isoenzymes expressed in different vertebrate species is variable. The two extremes are represented by humans, which express a single enzyme (AOX1) in many organs and mice or rats which are characterized by tissue-specific expression of four isoforms (AOX1, AOX2, AOX3, and AOX4). In vertebrates each AOX isoenzyme is the product of a distinct gene consisting of 35 highly conserved exons. The extant species-specific complement of AOX isoenzymes is the result of a complex evolutionary process consisting of a first phase characterized by a series of asynchronous gene duplications and a second phase where the pseudogenization and gene deletion events prevail. In the last few years remarkable advances in the elucidation of the structural characteristics and the catalytic mechanisms of mammalian AOXs have been made thanks to the successful crystallization of human AOX1 and mouse AOX3. Much less is known about the physiological function and physiological substrates of human AOX1 and other mammalian AOX isoenzymes, although the importance of these proteins in xenobiotic metabolism is fairly well established and their relevance in drug development is increasing. This review article provides an overview and a discussion of the current knowledge on mammalian AOX.

}, issn = {1432-0738}, doi = {10.1007/s00204-016-1683-1}, url = {http://dx.doi.org/10.1007/s00204-016-1683-1}, author = {Terao, Mineko and Rom{\~a}o, Maria Jo{\~a}o and Leimk{\"u}hler, Silke and Bolis, Marco and Fratelli, Maddalena and Coelho, Catarina and Santos-Silva, Teresa and Garattini, Enrico} } @article {PMID:26287794, title = {Ligand Binding to Chlorite Dismutase from Magnetospirillum sp}, journal = {The journal of physical chemistry. B}, volume = {119}, number = {43}, year = {2015}, note = {n/a}, month = {October}, pages = {13859{\textemdash}13869}, abstract = {n/a}, issn = {1520-6106}, doi = {10.1021/acs.jpcb.5b04141}, url = {http://dx.doi.org/10.1021/acs.jpcb.5b04141}, author = {De Schutter, Amy and Correia, Hugo D. and Freire, Diana M and Rivas, Mar{\'\i}a G and Rizzi, Alberto and Santos-Silva, Teresa and Gonz{\'a}lez, Pablo J and Van Doorslaer, Sabine} } @article {10239, title = {Structural and mechanistic insights on nitrate reductases}, journal = {Protein Science}, volume = {24}, year = {2015}, note = {

ISI Document Delivery No.: DB1TNCoelho, Catarina Romao, Maria Joao
Life Sciences (LS)
Wiley-blackwell
Hoboken

}, month = {2015}, pages = {1901-1911}, abstract = {

Nitrate reductases (NR) belong to the DMSO reductase family of Mo-containing enzymes and perform key roles in the metabolism of the nitrogen cycle, reducing nitrate to nitrite. Due to variable cell location, structure and function, they have been divided into periplasmic (Nap), cytoplasmic, and membrane-bound (Nar) nitrate reductases. The first crystal structure obtained for a NR was that of the monomeric NapA from Desulfovibrio desulfuricans in 1999. Since then several new crystal structures were solved providing novel insights that led to the revision of the commonly accepted reaction mechanism for periplasmic nitrate reductases. The two crystal structures available for the NarGHI protein are from the same organism (Escherichia coli) and the combination with electrochemical and spectroscopic studies also lead to the proposal of a reaction mechanism for this group of enzymes. Here we present an overview on the current advances in structural and functional aspects of bacterial nitrate reductases, focusing on the mechanistic implications drawn from the crystallographic data.

}, keywords = {Biochemistry \& Molecular Biology, Biophysics, dehydrogenase, molybdenum enzymes, haemophilus-influenzae, catalytic, diheme cytochrome-c, crystal-structure, escherichia-coli, formate, enzymatic mechanism, marinobacter-hydrocarbonoclasticus 617, dimethyl-sulfoxide reductase,, mechanism, angstrom resolution, molybdenum enzymes, nitrogen cycle, DMSO reductase family, periplasmic, nitrate reductase, membrane-bound nitrate reductase, crystal structure,}, isbn = {0961-8368}, url = {://CCC:000368292000001http://www.wiley.com/WileyCDA/}, author = {Coelho, C. and Romao, M. J.} } @article {10238, title = {Structural insights into xenobiotic and inhibitor binding to human aldehyde oxidase}, journal = {Nat Chem Biol}, volume = {11}, year = {2015}, note = {n/a}, month = {2015}, pages = {779-83}, isbn = {1552-4469}, url = {http://www.ncbi.nlm.nih.gov/pubmed/26322824}, author = {Coelho, C. and Foti, A. and Hartmann, T. and Santos-Silva, T. and Leimk and Rom} } @article {9429, title = {Aromatic aldehydes at the active site of aldehyde oxidoreductase from Desulfovibrio gigas: reactivity and molecular details of the enzyme-substrate and enzyme-product interaction}, journal = {Journal of Biological Inorganic Chemistry}, volume = {20}, number = {2}, year = {2015}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013 Romao, Maria/0000-0002-3004-0543 0

}, pages = {219-229}, type = {Journal Article}, abstract = {

Desulfovibrio gigas aldehyde oxidoreductase (DgAOR) is a mononuclear molybdenum-containing enzyme from the xanthine oxidase (XO) family, a group of enzymes capable of catalyzing the oxidative hydroxylation of aldehydes and heterocyclic compounds. The kinetic studies reported in this work showed that DgAOR catalyzes the oxidative hydroxylation of aromatic aldehydes, but not heterocyclic compounds. NMR spectroscopy studies using C-13-labeled benzaldehyde confirmed that DgAOR catalyzes the conversion of aldehydes to the respective carboxylic acids. Steady-state kinetics in solution showed that high concentrations of the aromatic aldehydes produce substrate inhibition and in the case of 3-phenyl propionaldehyde a suicide substrate behavior. Hydroxyl-substituted aromatic aldehydes present none of these behaviors but the kinetic parameters are largely affected by the position of the OH group. High-resolution crystallographic structures obtained from single crystals of active-DgAOR soaked with benzaldehyde showed that the side chains of Phe(425) and Tyr(535) are important for the stabilization of the substrate in the active site. On the other hand, the X-ray data of DgAOR soaked with trans-cinnamaldehyde showed a cinnamic acid molecule in the substrate channel. The X-ray data of DgAOR soaked with 3-phenyl propionaldehyde showed clearly how high substrate concentrations inactivate the enzyme by binding covalently at the surface of the enzyme and blocking the substrate channel. The different reactivity of DgAOR versus aldehyde oxidase and XO towards aromatic aldehydes and N-heterocyclic compounds is explained on the basis of the present kinetic and structural data.

}, issn = {0949-8257}, doi = {10.1007/s00775-014-1196-4}, url = {://WOS:000350236100005}, author = {Correia, Hugo D. and Marangon, Jacopo and Brondino, Carlos D. and Moura, Jose J. G. and Romao, Maria J. and Gonzalez, Pablo J. and Santos-Silva, Teresa} } @article {9430, title = {A contribution to the rational design of Ru(CO)(3)Cl2L complexes for in vivo delivery of CO}, journal = {Dalton Transactions}, volume = {44}, number = {11}, year = {2015}, note = {

Times Cited: 1 Romao, Maria/A-4115-2013 Romao, Maria/0000-0002-3004-0543 1

}, pages = {5058-5075}, type = {Journal Article}, abstract = {

A few ruthenium based metal carbonyl complexes, e.g. CORM-2 and CORM-3, have therapeutic activity attributed to their ability to deliver CO to biological targets. In this work, a series of related complexes with the formula [Ru(CO)(3)Cl2L] (L = DMSO (3), L-H3CSO(CH2)(2)CH(NH2)CO2H) (6a); D,L-H3CSO(CH2)(2)CH-(NH2)CO2H (6b); 3-NC5H4(CH2)(2)SO3.Na (7); 4-NC5H4(CH2)(2)SO3Na (8); PTA (9); DAPTA (10); H3CS-(CH2)(2)CH(OH) CO2H (11); CNCMe2CO2Me (12); CNCMeEtCO2Me (13); CN(c-C3H4)CO2Et) (14)) were designed, synthesized and studied. The effects of L on their stability, CO release profile, cytotoxicity and anti-inflammatory properties are described. The stability in aqueous solution depends on the nature of L as shown using HPLC and LC-MS studies. The isocyanide derivatives are the least stable complexes, and the S-bound methionine oxide derivative is the more stable one. The complexes do not release CO gas to the headspace, but release CO2 instead. X-ray diffraction of crystals of the model protein Hen Egg White Lysozyme soaked with 6b (4UWN) and 8 (4UWV) shows the addition of Ru-II(CO)(H2O)(4) at the His15 binding site. Soakings with 7 (4UWU) produced the metallacarboxylate [Ru(COOH)(CO)(H2O)(3)](+) bound to the His15 site. The aqueous chemistry of these complexes is governed by the water-gas shift reaction initiated with the nucleophilic attack of HO- on coordinated CO. DFT calculations show this addition to be essentially barrierless. The complexes have low cytotoxicity and low hemolytic indices. Following i.v. administration of CORM-3, the in vivo bio-distribution of CO differs from that obtained with CO inhalation or with heme oxygenase stimulation. A mechanism for CO transport and delivery from these complexes is proposed.

}, issn = {1477-9226}, doi = {10.1039/c4dt02966f}, url = {://WOS:000350488300027}, author = {Seixas, Joao D. and Santos, Marino F. A. and Mukhopadhyay, Abhik and Coelho, Ana C. and Reis, Patricia M. and Veiros, Luis F. and Marques, Ana R. and Penacho, Nuno and Goncalves, Ana M. L. and Romao, Maria J. and Bernardes, Goncalo J. L. and Santos-Silva, Teresa and Romao, Carlos C.} } @article {9428, title = {Insights into the structural determinants of substrate specificity and activity in mouse aldehyde oxidases}, journal = {Journal of Biological Inorganic Chemistry}, volume = {20}, number = {2}, year = {2015}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Coelho, Catarina /D-2073-2013 Romao, Maria/0000-0002-3004-0543; Coelho, Catarina /0000-0003-1852-5406 0

}, pages = {209-217}, type = {Journal Article}, abstract = {

In this work, a combination of homology modeling and molecular dynamics (MD) simulations was used to investigate the factors that modulate substrate specificity and activity of the mouse AOX isoforms: mAOX1, mAOX2 (previously mAOX3l1), mAOX3 and mAOX4. The results indicate that the AOX isoform structures are highly preserved and even more conserved than the corresponding amino acid sequences. The only differences are at the protein surface and substrate-binding site region. The substrate-binding site of all isoforms consists of two regions: the active site, which is highly conserved among all isoforms, and a isoform-specific region located above. We predict that mAOX1 accepts a broader range of substrates of different shape, size and nature relative to the other isoforms. In contrast, mAOX4 appears to accept a more restricted range of substrates. Its narrow and hydrophobic binding site indicates that it only accepts small hydrophobic substrates. Although mAOX2 and mAOX3 are very similar to each other, we propose the following pairs of overlapping substrate specificities: mAOX2/mAOX4 and mAOX3/mAXO1. Based on these considerations, we propose that the catalytic activity between all isoforms should be similar but the differences observed in the binding site might influence the substrate specificity of each enzyme. These results also suggest that the presence of several AOX isoforms in mouse allows them to oxidize more efficiently a wider range of substrates. This contrasts with the same or other organisms that only express one isoform and are less efficient or incapable of oxidizing the same type of substrates.

}, issn = {0949-8257}, doi = {10.1007/s00775-014-1198-2}, url = {://WOS:000350236100004}, author = {Cerqueira, Nuno M. F. S. A. and Coelho, Catarina and Bras, Natercia F. and Fernandes, Pedro A. and Garattini, Enrico and Terao, Mineko and Romao, Maria Joao and Ramos, Maria Joao} } @article {9898, title = {Ionic-Liquid-Functionalized Mineral Particles for Protein Crystallization}, journal = {Crystal Growth \& Design}, volume = {15}, number = {6}, year = {2015}, note = {

ISI Document Delivery No.: CK0JB Times Cited: 0 Cited Reference Count: 42 Kowacz, M. Marchel, M. Juknaite, L. Esperanca, J. M. S. S. Romao, M. J. Carvalho, A. L. Rebelo, L. P. N. Esperanca, Jose/B-5116-2008; Esperanca, Jose/0000-0001-9615-8678; Marchel, Mateusz/0000-0002-2274-1139; Juknaite, Lina/0000-0002-5739-7788 Fundacao para a Ciencia e a Tecnologia (FCT), Portugal [PTDC/BBB-BEP/3058/2012, PEst-OE/EQB/LA0004/2013, UID/Multi/04378/2013, PEst-C/EQB/LA0006/2013, RECI/BBB-BEP/0124/2012, SFRH/BPD/63554/2009]; Investigator FCT Program; COMPETE Program The authors thank Dr. Teresa Santos-Silva and Ph.D. student Hugo Correia for kindly providing the crystallization conditions for bovine blood hemoglobin and pancreatic trypsin. The authors acknowledge financial support from Fundacao para a Ciencia e a Tecnologia (FCT), Portugal, through R\&D Projects PTDC/BBB-BEP/3058/2012, PEst-OE/EQB/LA0004/2013 (to ITQB), UID/Multi/04378/2013, PEst-C/EQB/LA0006/2013 (to Associate Lab UCIBIO-REQUIMTE), and RECI/BBB-BEP/0124/2012 and also through a postdoctoral grant (SFRH/BPD/63554/2009) and a contract under the Investigator FCT 2012 Program and the COMPETE Program. The authors also thank the ESRF (Grenoble, France) and the Diamond Light Source (Didcot, U.K.) for access to data collection facilities. 9 Amer chemical soc Washington 1528-7505

}, pages = {2994-3003}, type = {Journal Article}, abstract = {

Nucleation is a critical step determining the outcome of the entire crystallization process. Finding an effective nucleant for protein crystallization is of utmost importance for structural biology. The latter relies on good-quality crystals to solve the three-dimensional structures of macromolecules. In this study we show that crystalline barium sulfate (BaSO4) with an etched and/or ionic liquid (IL)-functionalized surface (1) can induce protein nucleation at concentrations well below the concentration needed to promote crystal growth under control conditions, (2) can shorten the nucleation time, (3) can increase the growth rate, and finally (4) may help to improve the protein crystal morphology. These effects were shown for lysozyme, RNase A, trypsin, proteinase K, myoglobin, and hemoglobin. Therefore, the use of BaSO4 particles enables us to reduce the amount of protein in crystallization trials and increases the chance of obtaining protein crystals of the desired quality. In the context of the underlying mechanism, it is shown that the protein-solid contact formation is governed by the interaction of the polar compartments of the biomacromolecule with the support. The tendency of a protein to concentrate near the solid surface is enhanced by both the hydrophobicity of the protein and that of the surface (tuned by the functionalizing IL). These mechanisms of interaction of biomacromolecules with inorganic hydrophilic solids correspond to the principles of amphiphilic IL-mineral interactions.

}, keywords = {heterogeneous nucleation crystal nucleation adsorption stability surfaces water lysozyme fluorescence polystyrene suspensions Chemistry Crystallography Materials Science}, issn = {1528-7483}, doi = {10.1021/acs.cgd.5b00403}, url = {://WOS:000355890400056}, author = {Kowacz, M. and Marchel, M. and Juknaite, L. and Esperanca, Jmss and Romao, M. J. and Carvalho, A. L. and Rebelo, L. P. N.} } @article {11737, title = {TupA: A Tungstate Binding Protein in the Periplasm of Desulfovibrio alaskensis G20}, journal = {International Journal of Molecular Sciences}, volume = {15}, year = {2014}, note = {n/a}, month = {2014/05/29/accep}, pages = {11783 - 11798}, publisher = {MDPI}, abstract = {

The TupABC system is involved in the cellular uptake of tungsten and belongs to the ABC (ATP binding cassette)-type transporter systems. The TupA component is a periplasmic protein that binds tungstate anions, which are then transported through the membrane by the TupB component using ATP hydrolysis as the energy source (the reaction catalyzed by the ModC component). We report the heterologous expression, purification, determination of affinity binding constants and crystallization of the Desulfovibrio alaskensis G20 TupA. The tupA gene (locus tag Dde_0234) was cloned in the pET46 Enterokinase/Ligation-Independent Cloning (LIC) expression vector, and the construct was used to transform BL21 (DE3) cells. TupA expression and purification were optimized to a final yield of 10 mg of soluble pure protein per liter of culture medium. Native polyacrylamide gel electrophoresis was carried out showing that TupA binds both tungstate and molybdate ions and has no significant interaction with sulfate, phosphate or perchlorate. Quantitative analysis of metal binding by isothermal titration calorimetry was in agreement with these results, but in addition, shows that TupA has higher affinity to tungstate than molybdate. The protein crystallizes in the presence of 30\% (w/v) polyethylene glycol 3350 using the hanging-drop vapor diffusion method. The crystals diffract X-rays beyond 1.4 {\r A} resolution and belong to the P2(1) space group, with cell parameters a = 52.25 {\r A}, b = 42.50 {\r A}, c = 54.71 {\r A}, β = 95.43{\textdegree}. A molecular replacement solution was found, and the structure is currently under refinement.

}, isbn = {1422-0067}, url = {http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4139814/}, author = {Otrelo-Cardoso, Ana Rita and Nair, Rashmi R and Correia, M{\'a}rcia A. S. and Rivas, Maria G. and Santos-Silva, Teresa} } @article {7423, title = {Biochemical, Stabilization and Crystallization Studies on a Molecular Chaperone (PaoD) Involved in the Maturation of Molybdoenzymes}, journal = {Plos One}, volume = {9}, number = {1}, year = {2014}, note = {

Times Cited: 0 Cabrita, Eurico/E-8053-2011 Cabrita, Eurico/0000-0002-0720-2751 0

}, type = {Journal Article}, abstract = {n/a}, issn = {1932-6203}, doi = {10.1371/journal.pone.0087295}, url = {://WOS:000330621900083}, author = {Otrelo-Cardoso, Ana Rita and Schwuchow, Viola and Rodrigues, David and Cabrita, Eurico J. and Leimkuehler, Silke and Romao, Maria Joao and Santos-Silva, Teresa} } @article {CostaPessoa201460, title = {New insights on vanadium binding to human serum transferrin}, journal = {Inorganica Chimica Acta}, volume = {420}, year = {2014}, note = {

Recent Advances in Vanadium Chemistry Special Issue

}, pages = {60 - 68}, abstract = {

Abstract The knowledge on the binding of vanadium ions and complexes to serum proteins and how vanadium might be transported in blood and up-taken by cells has received much attention during the last decade, particularly as far as the transport of VIVO2+ is concerned. In this work we revise and discuss some relevant aspects of previous research, namely the two main types of binding proposed for transport of VIVO(carrier)2 complexes. New results, obtained by circular dichroism (CD), \{EPR\} and gel electrophoresis, regarding the binding of vanadium to hTF in the oxidation states +5 and +3 are also presented. Namely, evidences for the binding of VV-species to diferric-transferrin, designated by (FeIII)2hTF, as well as to (AlIII)2hTF, are presented and discussed, the possibility of up-take of vanadate by cells through (FeIII)2hTF endocytosis being suggested. It is also confirmed that \{VIII\} binds strongly to hTF, forming di-vanadium(III)-transferrin, designated by (VIII)2hTF, and gel electrophoresis experiments indicate that (VIII)2hTF corresponds to a {\textquoteleft}closed conformation{\textquoteright} similar to (FeIII)2hTF.

}, keywords = {Vanadium(III)-transferrin}, issn = {0020-1693}, doi = {http://dx.doi.org/10.1016/j.ica.2013.11.025}, url = {http://www.sciencedirect.com/science/article/pii/S0020169313006373}, author = {Jo{\~a}o Costa Pessoa and Gisela Gon{\c c}alves and Roy, Somnath and Isabel Correia and Mehtab, Sameena and Santos, Marino F. A. and Santos-Silva, Teresa} } @article {7422, title = {Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis}, journal = {International Journal of Molecular Sciences}, volume = {15}, number = {2}, year = {2014}, note = {

Times Cited: 0 0

}, pages = {2223-2236}, type = {Journal Article}, abstract = {n/a}, issn = {1422-0067}, doi = {10.3390/ijms15022223}, url = {://WOS:000335776400032}, author = {Otrelo-Cardoso, Ana Rita and da Silva Correia, Marcia Alexandra and Schwuchow, Viola and Svergun, Dmitri I. and Romao, Maria Joao and Leimkuehler, Silke and Santos-Silva, Teresa} } @article {9900, title = {Use of Gold Nanoparticles as Additives in Protein Crystallization}, journal = {Crystal Growth \& Design}, volume = {14}, number = {1}, year = {2014}, note = {

ISI Document Delivery No.: 284RC Times Cited: 0 Cited Reference Count: 19 Ribeiro, Diana Kulakova, Alina Quaresma, Pedro Pereira, Eulalia Bonifacio, Cecilia Romao, Maria Joao Franco, Ricardo Carvalho, Ana Luisa Portuguese Science and Technology Foundation (FCT-MEC); COMPETE [PTDC/CTM-NAN/112241/2009, PEst-C/EQB/LA0006/2011, PEst-C/EQB/LA0006/2013] The authors would like to acknowledge Abhik Mukhopadhyay for assistance in data collection and processing of lysozyme crystals, Teresa Santos-Silva, Angelina Palma, Benedita Pinheiro, Marcia Correia, and Catarina Coelho for kindly providing the model proteins used as "real" cases in this study, and Marino Santos for data processing from albumin crystals and assistance in preparation of figures. We also acknowledge assistance in synchrotron data collection at ESRF (Grenoble, France) and SLS (Villigen, Switzerland). The work was supported by the Portuguese Science and Technology Foundation (FCT-MEC) and COMPETE through Grants PTDC/CTM-NAN/112241/2009 to R.F. and Grants PEst-C/EQB/LA0006/2011 and PEst-C/EQB/LA0006/2013 (to Associate Lab REQUIMTE). Amer chemical soc Washington

}, pages = {222-227}, type = {Journal Article}, abstract = {

Gold nanoparticles (AuNPs) exhibit unique properties that have made them a very attractive material for application in biological assays. Given the potentially interesting interactions between AuNPs and biological macromolecules, we investigated AuNPs-induced protein crystal growth. Differently functionalized AuNPs were tested as additives in cocrystallization studies with model proteins (hen egg white lysozyme (HEWL), ribonuclease A (RNase A), and proteinase K) as well as with case studies where there were problems in obtaining well-diffracting crystals. Trials were performed considering different crystallization drawbacks, from total absence of crystals to improvement of crystal morphology, size, twinning, and number of crystals per drop. Improvement of some of these factors was observed in the cases of HEWL, RNase A, phenylalanine hydroxylase (PAR), myoglobin, native aldehyde oxidase (AOH), and human albumin. In these proteins, the presence of the AuNPs promoted an increase in the size and/or better crystal morphology. From the systematic trials and subsequent observations, it can be concluded that the introduction of AuNPs should definitely be considered in crystal optimization trials to improve previously determined crystallization conditions.

}, keywords = {Chemistry Crystallography Materials Science}, issn = {1528-7483}, doi = {10.1021/cg4014398}, url = {://WOS:000329337000029}, author = {Ribeiro, D and Kulakova, A. and P. Quaresma and Pereira, E. and Bonifacio, C. and Romao, M. J. and Franco, R. and Carvalho, A. L.} } @article {7292, title = {Characterization of a versatile organometallic pro-drug (CORM) for experimental CO based therapeutics}, journal = {Dalton Transactions}, volume = {42}, number = {17}, year = {2013}, note = {

Times Cited: 4 Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Reis, Patricia/I-7422-2013; Coelho, Ana Catarina/J-3331-2013; Romao, Maria/A-4115-2013; Santos-Silva, Teresa/D-2050-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013; PTMS, RNEM/C-1589-2014 Reis, Patricia/0000-0001-6320-7496; Romao, Maria/0000-0002-3004-0543; 5

}, pages = {5985-5998}, type = {Journal Article}, abstract = {n/a}, issn = {1477-9226}, doi = {10.1039/c2dt32174b}, url = {://WOS:000317013000005}, author = {Seixas, Joao D. and Mukhopadhyay, Abhik and Santos-Silva, Teresa and Otterbein, Leo E. and Gallo, David J. and Rodrigues, Sandra S. and Guerreiro, Bruno H. and Goncalves, Ana M. L. and Penacho, Nuno and Marques, Ana R. and Coelho, Ana C. and Reis, Patricia M. and Romao, Maria J. and Romao, Carlos C.} } @article {7421, title = {Determinants of selective group reduction in the TNT-bound xenobiotic reductase B from P. putida}, journal = {European Biophysics Journal with Biophysics Letters}, volume = {42}, year = {2013}, note = {

Times Cited: 0 1 9th European-Biophysical-Societies-Association Congress Jul 13-17, 2013 Lisbon, PORTUGAL European Biophys Soc Assoc; Soc Portuguesa Biofisica 0

}, pages = {S179-S179}, type = {Journal Article}, abstract = {n/a}, issn = {0175-7571; 1432-1017}, url = {://WOS:000330215300550}, author = {Mukhopadhyay, A. and Bursakov, S. A. and Ramos, J. L. and Wittich, R. M. and Kladova, A. V. and Romao, M. J. and van Dillewijn, P. and Carvalho, A. L.} } @article {7419, title = {Identification of Crucial Amino Acids in Mouse Aldehyde Oxidase 3 That Determine Substrate Specificity}, journal = {Plos One}, volume = {8}, number = {12}, year = {2013}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013 Romao, Maria/0000-0002-3004-0543 0

}, type = {Journal Article}, abstract = {n/a}, issn = {1932-6203}, doi = {10.1371/journal.pone.0082285}, url = {://WOS:000328735700053}, author = {Mahro, Martin and Bras, Natercia F. and Cerqueira, Nuno M. F. S. A. and Teutloff, Christian and Coelho, Catarina and Romao, Maria Joao and Leimkuehler, Silke} } @article {7202, title = {Induced peroxidase activity of haem containing nitrate reductases revealed by protein film electrochemistry}, journal = {Journal of Electroanalytical Chemistry}, volume = {693}, year = {2013}, note = {

Times Cited: 2 Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Romao, Maria/A-4115-2013; dos Santos, Margarida/H-7897-2012; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; dos Santos, Margarida/0000-0001-7531-757X; 2

}, pages = {105-113}, type = {Journal Article}, abstract = {n/a}, issn = {1572-6657}, doi = {10.1016/j.jelechem.2013.01.030}, url = {://WOS:000317445800016}, author = {Coelho, Catarina and Marangon, Jacopo and Rodrigues, David and Moura, Jose J. G. and Romao, Maria Joao and Paes de Sousa, Patricia M. and Correia dos Santos, Margarida M.} } @article {7243, title = {Interaction of vanadium(IV) with human serum apo-transferrin}, journal = {Journal of Inorganic Biochemistry}, volume = {121}, year = {2013}, note = {

Times Cited: 7 Romao, Maria/A-4115-2013; Roy, Dr. Somnath/F-6257-2012; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Tomaz, Ana Isabel/D-6988-2012; Santos-Silva, Teresa/D-2050-2013; REQUIMTE, SMB/M-5694-2013; Costa Pessoa, Joao/H-9078-2012; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Roy, Dr. Somnath/0000-0002-0031-606X; Tomaz, Ana Isabel/0000-0002-2249-4684; Costa Pessoa, Joao/0000-0002-3978-9964; 7

}, pages = {187-195}, type = {Journal Article}, abstract = {n/a}, issn = {0162-0134}, doi = {10.1016/j.jinorgbio.2012.12.020}, url = {://WOS:000316305700022}, author = {Mehtab, Sameena and Goncalves, Gisela and Roy, Somnath and Tomaz, Ana Isabel and Santos-Silva, Teresa and Santos, Marino F. A. and Romao, Maria J. and Jakusch, Tamas and Kiss, Tamas and Pessoa, Joao Costa} } @article {7420, title = {Kinetic and Structural Studies of Aldehyde Oxidoreductase from Desulfovibrio gigas Reveal a Dithiolene-Based Chemistry for Enzyme Activation and Inhibition by H2O2}, journal = {Plos One}, volume = {8}, number = {12}, year = {2013}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013 Romao, Maria/0000-0002-3004-0543 0

}, type = {Journal Article}, abstract = {n/a}, issn = {1932-6203}, doi = {10.1371/journal.pone.0083234}, url = {://WOS:000329325200045}, author = {Marangon, Jacopo and Correia, Hugo D. and Brondino, Carlos D. and Moura, Jose J. G. and Romao, Maria J. and Gonzalez, Pablo J. and Santos-Silva, Teresa} } @article {7429, title = {Overexpression, crystallization and preliminary X-ray crystallographic analysis of glucuronoxylan xylanohydrolase (Xyn30A) from Clostridium thermocellum}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {69}, year = {2013}, note = {

Times Cited: 0 Prates, Jose/K-9934-2013; Romao, Maria/A-4115-2013 Prates, Jose/0000-0003-1032-5987; Romao, Maria/0000-0002-3004-0543 12 0

}, pages = {1440-1442}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309113025050}, url = {://WOS:000327942200030}, author = {Verma, Anil Kumar and Goyal, Arun and Freire, Filipe and Bule, Pedro and Venditto, Immacolata and Bras, Joana L. A. and Santos, Helena and Cardoso, Vania and Bonifacio, Cecilia and Thompson, Andrew and Romao, Maria Joao and Prates, Jose A. M. and Ferreira, Luis M. A. and Fontes, Carlos M. G. A. and Najmudin, Shabir} } @article {7302, title = {Solution structure, dynamics and binding studies of a family 11 carbohydrate-binding module from Clostridium thermocellum (CtCBM11)}, journal = {Biochemical Journal}, volume = {451}, year = {2013}, note = {

Times Cited: 1 Viegas, Aldino/A-1889-2011; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Romao, Maria/A-4115-2013; Cabrita, Eurico/E-8053-2011; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013; Caparica, PTNMR/E-5112-2013 Viegas, Aldino/0000-0003-1733-136X; Romao, Maria/0000-0002-3004-0543; Cabrita, Eurico/0000-0002-0720-2751; Carvalho, Ana Luisa/0000-0002-3824-0240; 2 1

}, pages = {289-300}, type = {Journal Article}, abstract = {n/a}, issn = {0264-6021}, doi = {10.1042/bj20120627}, url = {://WOS:000317443500016}, author = {Viegas, Aldino and Sardinha, Joao and Freire, Filipe and Duarte, Daniel F. and Carvalho, Ana L. and Fontes, Carlos M. G. A. and Romao, Maria J. and Macedo, Anjos L. and Cabrita, Eurico J.} } @article {7424, title = {The Structural Basis of the Recognition of Di-glucosylated N-glycans by the ER Lectin Malectin}, journal = {Glycobiology}, volume = {23}, number = {11}, year = {2013}, note = {

Times Cited: 0 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543 Annual Conference of the Society-for-Glycobiology Nov 17-20, 2013 St Petersburg, FL Soc Glycobiol 0

}, pages = {1368-1369}, type = {Journal Article}, abstract = {n/a}, issn = {0959-6658; 1460-2423}, url = {://WOS:000326972400123}, author = {Palma, Angelina S. and Pinheiro, Benedita and Liu, Yan and Takeda, Yoichi and Chai, Wengang and Ito, Yukishige and Romao, Maria Joao and Carvalho, Ana Luisa and Feizi, Ten} } @article {7425, title = {Study of the cohesin-dockerin interaction and its role in the C. thermocellum cellulosome assembly}, journal = {European Biophysics Journal with Biophysics Letters}, volume = {42}, year = {2013}, note = {

Times Cited: 0 1 9th European-Biophysical-Societies-Association Congress Jul 13-17, 2013 Lisbon, PORTUGAL European Biophys Soc Assoc; Soc Portuguesa Biofisica 0

}, pages = {S180-S180}, type = {Journal Article}, abstract = {n/a}, issn = {0175-7571; 1432-1017}, url = {://WOS:000330215300555}, author = {Pinheiro, B. A. and Carvalho, A. L. and Romao, M. J. and Fontes, C. M.} } @article {7427, title = {Unraveling new functions and modes of action of molybdenum-dependent enzymes}, journal = {European Biophysics Journal with Biophysics Letters}, volume = {42}, year = {2013}, note = {

Times Cited: 0 1 9th European-Biophysical-Societies-Association Congress Jul 13-17, 2013 Lisbon, PORTUGAL European Biophys Soc Assoc; Soc Portuguesa Biofisica 0

}, pages = {S35-S35}, type = {Journal Article}, abstract = {n/a}, issn = {0175-7571; 1432-1017}, url = {://WOS:000330215300004}, author = {Romao, M. J.} } @article {7428, title = {Vanadium compounds as prospective therapeutics: X-ray structure of protein adducts}, journal = {European Biophysics Journal with Biophysics Letters}, volume = {42}, year = {2013}, note = {

Times Cited: 0 1 9th European-Biophysical-Societies-Association Congress Jul 13-17, 2013 Lisbon, PORTUGAL European Biophys Soc Assoc; Soc Portuguesa Biofisica 0

}, pages = {S181-S181}, type = {Journal Article}, abstract = {n/a}, issn = {0175-7571; 1432-1017}, url = {://WOS:000330215300560}, author = {Santos, M. F. A. and Oliveira, A. R. and Somnath, R. and Romao, M. J. and Pessoa, J. C. and Santos-Silva, T.} } @inbook {7183, title = {ESCHERICHIA COLI EXPRESSION, PURIFICATION, CRYSTALLIZATION, AND STRUCTURE DETERMINATION OF BACTERIAL COHESIN-DOCKERIN COMPLEXES}, booktitle = {Cellulases}, series = {Methods in Enzymology}, volume = {510}, year = {2012}, note = {

Times Cited: 2 Romao, Maria/A-4115-2013; Viegas, Aldino/A-1889-2011; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Prates, Jose/K-9934-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Viegas, Aldino/0000-0003-1733-136X; Prates, Jose/0000-0003-1032-5987; Carvalho, Ana Luisa/0000-0002-3824-0240; 2

}, pages = {395-415}, type = {Book Section}, abstract = {n/a}, isbn = {0076-6879 978-0-12-415931-0}, doi = {10.1016/b978-0-12-415931-0.00021-5}, url = {://WOS:000308194700021}, author = {Bras, Joana L. A. and Carvalho, Ana Luisa and Viegas, Aldino and Najmudin, Shabir and Alves, Victor D. and Prates, Jose A. M. and Ferreira, Luis M. A. and Romao, Maria J. and Gilbert, Harry J. and Fontes, Carlos M. G. A.}, editor = {Gilbert, H. J.} } @article {7201, title = {The First Mammalian Aldehyde Oxidase Crystal Structure INSIGHTS INTO SUBSTRATE SPECIFICITY}, journal = {Journal of Biological Chemistry}, volume = {287}, number = {48}, year = {2012}, note = {

Times Cited: 9 Romao, Maria/A-4115-2013; Coelho, Catarina /D-2073-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Coelho, Catarina /0000-0003-1852-5406; 9

}, pages = {40690-40702}, type = {Journal Article}, abstract = {n/a}, issn = {0021-9258}, doi = {10.1074/jbc.M112.390419}, url = {://WOS:000311448800058}, author = {Coelho, Catarina and Mahro, Martin and Trincao, Jose and Carvalho, Alexandra T. P. and Ramos, Maria Joao and Terao, Mineko and Garattini, Enrico and Leimkuehler, Silke and Romao, Maria Joao} } @article {9901, title = {Hofmeister effects of ionic liquids in protein crystallization: Direct and water-mediated interactions}, journal = {Crystengcomm}, volume = {14}, number = {15}, year = {2012}, note = {

ISI Document Delivery No.: 968QN Times Cited: 14 Cited Reference Count: 69 Kowacz, Magdalena Mukhopadhyay, Abhik Carvalho, Ana Luisa Esperanca, Jose M. S. S. Romao, Maria J. Rebelo, Luis Paulo N. Rebelo, Luis Paulo/B-5285-2008; Romao, Maria/A-4115-2013; Mukhopadhyay, Abhik/D-2119-2013; Kowacz, Magdalena/D-2386-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Esperanca, Jose/B-5116-2008; Chaves, Pedro/K-1288-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Mukhopadhyay, Abhik/0000-0002-9808-7620; Rebelo, Luis Paulo/0000-0002-5247-2443; Romao, Maria/0000-0002-3004-0543; Mukhopadhyay, Abhik/0000-0002-7755-0401; Kowacz, Magdalena/0000-0001-5729-6816; Esperanca, Jose/0000-0001-9615-8678; Carvalho, Ana Luisa/0000-0002-3824-0240; Fundacao para a Ciencia e a Tecnologia, Portugal [PEst-C/EQB/LA0006/2011, PEst-OE/EQB/LA0004/2011, SFRH/BPD/30142/2006, SFRH/BPD/63554/2009]; Research Executive Agency [PERG05-GA-2009-249182] The authors acknowledge financial support from Fundacao para a Ciencia e a Tecnologia, Portugal, through grants PEst-C/EQB/LA0006/2011 to the Associate Lab REQUIMTE and PEst-OE/EQB/LA0004/2011 to the ITQB and through post-doctoral grants SFRH/BPD/30142/2006 and SFRH/BPD/63554/2009. The authors thank the Research Executive Agency for Marie Curie Reintegration grant PERG05-GA-2009-249182. The authors also acknowledge the SOLEIL synchrotron facility (Paris, France) for access and technical support during data collection. 14 9 60 Royal soc chemistry Cambridge

}, pages = {4912-4921}, type = {Journal Article}, abstract = {

We have performed experiments on the crystallization of two low molecular weight, positively charged proteins, lysozyme and ribonuclease A, using ionic liquids as either crystallization additives or, in particular cases, as precipitating agents. The ionic liquids (ILs) have been ordered according to their salting-in/out ability and the relative position of these ionic liquids in this ranking has been rationalized by considering their hydration properties (positive-negative, hydrophobic-hydrophilic). The ability to screen the effective charge of cationic proteins and aid protein nucleation (salting-out) has been shown to be superior for large polarizable anions with low charge density, negatively hydrated-Cl-, Br-, [SCN](-), methane-[C1SO3](-) and ethanesulfonates [C2SO3](-), than for anions with a relatively stable hydration shell, positively hydrated-lactate [Lac](-), butylsulfonate [C4SO3](-) and acetate [Ac](-). Upon increasing the background salt concentration, where electrostatic interactions are already effectively screened, the ability of the IL ions to stabilize proteins in solution (salting-in) has been shown to increase as the ions are likely to migrate to the non-polar protein surface and lower protein-water interfacial tension. This tendency is enhanced as the focus moves from those ions with positively hydrated hydrophilic compartments (e. g. [Ac](-)) to those with negatively hydrated groups (e. g. [C1SO3](-)) and the prevailing hydrophobic hydration (e. g. [C4SO3](-)). The observed inversion in the relative effect of ILs on protein crystallization with increasing ionic strength of the aqueous media has been interpreted as the differing effects of ion adsorption: charge screening and interfacial tension modification. Moreover, this work can further help in our understanding of the influence of ionic liquids on conformational changes of biomacromolecules in solution. Identification of the specific incorporation sites for choline and acetate ions, localized in two lysozyme crystals grown in pure IL solutions without any buffer or inorganic precipitant, can give us some insight into the role of the ionic liquid ions in protein structure development.

}, keywords = {surface-tension measurements molecular-dynamics simulations aqueous-solutions cytochrome-c salting-out functional conversion hydration lysozyme stability extraction Chemistry Crystallography}, issn = {1466-8033}, doi = {10.1039/c2ce25129a}, url = {://WOS:000305999500006}, author = {Kowacz, M. and Mukhopadhyay, A. and Carvalho, A. L. and Esperanca, Jmss and Romao, M. J. and Rebelo, L. P. N.} } @article {7283, title = {New insights into the chemistry of fac- Ru(CO)(3) (2+) fragments in biologically relevant conditions: The CO releasing activity of Ru(CO)(3)Cl-2(1,3-thiazole) , and the X-ray crystal structure of its adduct with lysozyme}, journal = {Journal of Inorganic Biochemistry}, volume = {117}, year = {2012}, note = {

Times Cited: 3 Romao, Carlos /D-1162-2010; Romao, Maria/A-4115-2013; Mukhopadhyay, Abhik/D-2119-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Reis, Patricia/I-7422-2013; Coelho, Ana Catarina/J-3331-2013; Chaves, Pedro/K-1288-2013; Santos-Silva, Teresa/D-2050-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013; PTMS, RNEM/C-1589-2014 Romao, Carlos /0000-0001-5061-3743; Romao, Maria/0000-0002-3004-0543; Mukhopadhyay, Abhik/0000-0002-7755-0401; Reis, Patricia/0000-0001-6320-7496; 3

}, pages = {285-291}, type = {Journal Article}, abstract = {n/a}, issn = {0162-0134}, doi = {10.1016/j.jinorgbio.2012.06.018}, url = {://WOS:000311819300034}, author = {Santos, M. F. A. and Seixas, J. D. and Coelho, A. C. and Mukhopadhyay, A. and Reis, P. M. and Romao, M. J. and Romao, C. C. and Santos-Silva, T.} } @article {7181, title = {Novel Clostridium thermocellum Type I Cohesin-Dockerin Complexes Reveal a Single Binding Mode}, journal = {Journal of Biological Chemistry}, volume = {287}, number = {53}, year = {2012}, note = {

Times Cited: 1 Romao, Maria/A-4115-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Prates, Jose/K-9934-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Prates, Jose/0000-0003-1032-5987; Carvalho, Ana Luisa/0000-0002-3824-0240; 2

}, pages = {44394-44405}, type = {Journal Article}, abstract = {n/a}, issn = {0021-9258}, doi = {10.1074/jbc.M112.407700}, url = {://WOS:000312938600036}, author = {Bras, Joana L. A. and Alves, Victor D. and Carvalho, Ana Luia and Najmudin, Shabir and Prates, Jose A. M. and Ferreira, Luis M. A. and Bolam, David N. and Romao, Maria Joao and Gilbert, Harry J. and Fontes, Carlos M. G. A.} } @article {7177, title = {Structural studies of the p22HBP/SOUL family of heme-binding proteins}, journal = {Febs Journal}, volume = {279}, year = {2012}, note = {

Times Cited: 0 Freire, Filipe/F-6505-2010; Romao, Maria/A-4115-2013; Macedo, Anjos/D-7216-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Freire, Filipe/0000-0002-5310-0668; Romao, Maria/0000-0002-3004-0543; Macedo, Anjos/0000-0002-2613-4838; Carvalho, Ana Luisa/0000-0002-3824-0240; Si 1 22nd IUBMB Congress/37th FEBS Congress Sep 04-09, 2012 Seville, SPAIN Iubmb; febs 0

}, pages = {458-458}, type = {Journal Article}, abstract = {n/a}, issn = {1742-464X}, url = {://WOS:000308128602495}, author = {Aveiro, S. S. and Freire, F. and Clayton, J. and Cameloc, M. and Carvalho, A. L. and Ferreira, G. C. and Romao, M. J. and Macedo, A. L. and Goodfellow, B. J.} } @article {7242, title = {Characterization and Crystallization of Mouse Aldehyde Oxidase 3: From Mouse Liver to Escherichia coli Heterologous Protein Expression}, journal = {Drug Metabolism and Disposition}, volume = {39}, number = {10}, year = {2011}, note = {

Times Cited: 5 Romao, Maria/A-4115-2013; Coelho, Catarina /D-2073-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Coelho, Catarina /0000-0003-1852-5406; 5

}, pages = {1939-1945}, type = {Journal Article}, abstract = {n/a}, issn = {0090-9556}, doi = {10.1124/dmd.111.040873}, url = {://WOS:000295026600021}, author = {Mahro, Martin and Coelho, Catarina and Trincao, Jose and Rodrigues, David and Terao, Mineko and Garattini, Enrico and Saggu, Miguel and Lendzian, Friedhelm and Hildebrandt, Peter and Romao, Maria Joao and Leimkuehler, Silke} } @article {7290, title = {CORM-3 Reactivity toward Proteins: The Crystal Structure of a Ru(II) Dicarbonyl-Lysozyme Complex}, journal = {Journal of the American Chemical Society}, volume = {133}, number = {5}, year = {2011}, note = {

Times Cited: 28 Romao, Carlos /D-1162-2010; Romao, Maria/A-4115-2013; Mukhopadhyay, Abhik/D-2119-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Carlos /0000-0001-5061-3743; Romao, Maria/0000-0002-3004-0543; Mukhopadhyay, Abhik/0000-0002-7755-0401; 29

}, pages = {1192-1195}, type = {Journal Article}, abstract = {n/a}, issn = {0002-7863}, doi = {10.1021/ja108820s}, url = {://WOS:000287228500014}, author = {Santos-Silva, Teresa and Mukhopadhyay, Abhik and Seixas, Joao D. and Bernardes, Goncalo J. L. and Romao, Carlos C. and Romao, Maria J.} } @article {7199, title = {The Crystal Structure of Cupriavidus necator Nitrate Reductase in Oxidized and Partially Reduced States}, journal = {Journal of Molecular Biology}, volume = {408}, number = {5}, year = {2011}, note = {

Times Cited: 18 Romao, Maria/A-4115-2013; Coelho, Catarina /D-2073-2013; Moura, Isabel/D-6339-2013; Moura, Jose/D-6426-2013; Gonzalez, Pablo/D-8200-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Coelho, Catarina /0000-0003-1852-5406; Moura, Isabel/0000-0003-0971-4977; 18

}, pages = {932-948}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1016/j.jmb.2011.03.016}, url = {://WOS:000291066200012}, author = {Coelho, Catarina and Gonzalez, Pablo J. and Moura, Jose J. G. and Moura, Isabel and Trincao, Jose and Romao, Maria Joao} } @article {7248, title = {Crystal structure of the zinc-, cobalt-, and iron-containing adenylate kinase from Desulfovibrio gigas: a novel metal-containing adenylate kinase from Gram-negative bacteria}, journal = {Journal of Biological Inorganic Chemistry}, volume = {16}, number = {1}, year = {2011}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Mukhopadhyay, Abhik/D-2119-2013; Moura, Isabel/D-6339-2013; Moura, Jose/D-6426-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Mukhopadhyay, Abhik/0000-0002-7755-0401; Moura, Isabel/0000-0003-0971-4977; 0

}, pages = {51-61}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s00775-010-0700-8}, url = {://WOS:000286789800008}, author = {Mukhopadhyay, A. and Kladova, A. V. and Bursakov, S. A. and Gavel, O. Yu and Calvete, J. J. and Shnyrov, V. L. and Moura, I. and Moura, J. J. G. and Romao, M. J. and Trincao, J.} } @article {7223, title = {Molecular Architecture and Structural Transitions of a Clostridium thermocellum Mini-Cellulosome}, journal = {Journal of Molecular Biology}, volume = {407}, number = {4}, year = {2011}, note = {

Times Cited: 7 Carvalho, Ana Luisa/G-5638-2011; Dias, Fernando/I-4861-2012; Romao, Maria/A-4115-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Dias, Fernando/0000-0001-8109-2063; Romao, Maria/0000-0002-3004-0543; Prates, Jose/0000-0003-1032-5987; 8

}, pages = {571-580}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1016/j.jmb.2011.01.060}, url = {://WOS:000288925200008}, author = {Garcia-Alvarez, Begona and Melero, Roberto and Dias, Fernando M. V. and Prates, Jose A. M. and Fontes, Carlos M. G. A. and Smith, Steven P. and Romao, Maria Joao and Carvalho, Ana Luisa and Llorca, Oscar} } @article {7241, title = {Overproduction, purification, crystallization and preliminary X-ray characterization of a novel carbohydrate-binding module of endoglucanase Cel5A from Eubacterium cellulosolvens}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {67}, year = {2011}, note = {

Times Cited: 2 Romao, Maria/A-4115-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Prates, Jose/0000-0003-1032-5987; 4 2

}, pages = {491-493}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309111004246}, url = {://WOS:000289738400018}, author = {Luis, Ana S. and Alves, Victor D. and Romao, Maria J. and Prates, Jose A. M. and Fontes, Carlos M. G. A. and Najmudin, Shabir} } @article {7184, title = {Purification, crystallization and preliminary X-ray characterization of the pentamodular arabinoxylanase CtXyl5A from Clostridium thermocellum}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {67}, year = {2011}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Correia, Marcia/D-2077-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Correia, Marcia/0000-0003-0636-8095; Prates, Jose/0000-0003-1032-5987; 7 0

}, pages = {833-836}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309111020823}, url = {://WOS:000293191400025}, author = {Bras, Joana L. A. and Correia, Marcia A. S. and Romao, Maria J. and Prates, Jose A. M. and Fontes, Carlos M. G. A. and Najmudin, Shabir} } @article {7182, title = {Structural insights into a unique cellulase fold and mechanism of cellulose hydrolysis}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {108}, number = {13}, year = {2011}, note = {

Times Cited: 16 Romao, Maria/A-4115-2013; Prates, Jose/K-9934-2013; Correia, Marcia/D-2077-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Prates, Jose/0000-0003-1032-5987; Correia, Marcia/0000-0003-0636-8095; Carvalho, Ana Luisa/0000-0002-3824-0240; 16

}, pages = {5237-5242}, type = {Journal Article}, abstract = {n/a}, issn = {0027-8424}, doi = {10.1073/pnas.1015006108}, url = {://WOS:000288894800024}, author = {Bras, Joana L. A. and Cartmell, Alan and Carvalho, Ana Luisa M. and Verze, Genny and Bayer, Edward A. and Vazana, Yael and Correia, Marcia A. S. and Prates, Jose A. M. and Ratnaparkhe, Supriya and Boraston, Alisdair B. and Romao, Maria J. and Fontes, Carlos M. G. A. and Gilbert, Harry J.} } @article {7285, title = {Towards Improved Therapeutic CORMs: Understanding the Reactivity of CORM-3 with Proteins}, journal = {Current Medicinal Chemistry}, volume = {18}, number = {22}, year = {2011}, note = {

Times Cited: 13 Romao, Carlos /D-1162-2010; Romao, Maria/A-4115-2013; Mukhopadhyay, Abhik/D-2119-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Carlos /0000-0001-5061-3743; Romao, Maria/0000-0002-3004-0543; Mukhopadhyay, Abhik/0000-0002-7755-0401; 13

}, pages = {3361-3366}, type = {Journal Article}, abstract = {n/a}, issn = {0929-8673}, url = {://WOS:000294405600010}, author = {Santos-Silva, T. and Mukhopadhyay, A. and Seixas, J. D. and Bernardes, G. J. L. and Romao, C. C. and Romao, M. J.} } @article {7253, title = {The 1.4 angstrom resolution structure of Paracoccus pantotrophus pseudoazurin}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {66}, year = {2010}, note = {

Times Cited: 6 Romao, Maria/A-4115-2013; Pauleta, Sofia/F-3619-2011; Moura, Isabel/D-6339-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Pauleta, Sofia/0000-0002-2149-9416; Moura, Isabel/0000-0003-0971-4977; 6

}, pages = {627-635}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309110013989}, url = {://WOS:000278165900002}, author = {Najmudin, Shabir and Pauleta, Sofia R. and Moura, Isabel and Romao, Maria J.} } @article {7266, title = {Family 42 carbohydrate-binding modules display multiple arabinoxylan-binding interfaces presenting different ligand affinities}, journal = {Biochimica Et Biophysica Acta-Proteins and Proteomics}, volume = {1804}, number = {10}, year = {2010}, note = {

Times Cited: 1 Dias, Fernando/I-4861-2012; Romao, Maria/A-4115-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Dias, Fernando/0000-0001-8109-2063; Romao, Maria/0000-0002-3004-0543; Prates, Jose/0000-0003-1032-5987; 1

}, pages = {2054-2062}, type = {Journal Article}, abstract = {n/a}, issn = {1570-9639}, doi = {10.1016/j.bbapap.2010.07.006}, url = {://WOS:000281920300012}, author = {Ribeiro, Teresa and Santos-Silva, Teresa and Alves, Victor D. and Dias, Fernando M. V. and Luis, Ana S. and Prates, Jose A. M. and Ferreira, Luis M. A. and Romao, Maria J. and Fontes, Carlos M. G. A.} } @article {7254, title = {Putting an N-terminal end to the Clostridium thermocellum xylanase Xyn10B story: Crystal structure of the CBM22-1-GH10 modules complexed with xylohexaose}, journal = {Journal of Structural Biology}, volume = {172}, number = {3}, year = {2010}, note = {

Times Cited: 9 Romao, Maria/A-4115-2013; Pinheiro, Benedita/D-2055-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Pinheiro, Benedita/0000-0002-3045-077X; Prates, Jose/0000-0003-1032-5987; 9

}, pages = {353-362}, type = {Journal Article}, abstract = {n/a}, issn = {1047-8477}, doi = {10.1016/j.jsb.2010.07.009}, url = {://WOS:000283964000018}, author = {Najmudin, Shabir and Pinheiro, Benedita A. and Prates, Jose A. M. and Gilbert, Harry J. and Romao, Maria J. and Fontes, Carlos M. G. A.} } @article {7203, title = {The use of ionic liquids as crystallization additives allowed to overcome nanodrop scaling up problems: A success case for producing diffraction-quality crystals of a nitrate reductase}, journal = {Journal of Crystal Growth}, volume = {312}, number = {5}, year = {2010}, note = {

Times Cited: 8 Romao, Maria/A-4115-2013; Coelho, Catarina /D-2073-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Coelho, Catarina /0000-0003-1852-5406; 8

}, pages = {714-719}, type = {Journal Article}, abstract = {n/a}, issn = {0022-0248}, doi = {10.1016/j.jcrysgro.2009.11.060}, url = {://WOS:000275137100016}, author = {Coelho, Catarina and Trincao, Jose and Romao, Maria Joao} } @article {7234, title = {Cobalt-, zinc- and iron-bound forms of adenylate kinase (AK) from the sulfate-reducing bacterium Desulfovibrio gigas: purification, crystallization and preliminary X-ray diffraction analysis}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {65}, year = {2009}, note = {

Times Cited: 1 Romao, Maria/A-4115-2013; Moura, Isabel/D-6339-2013; Moura, Jose/D-6426-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 1

}, pages = {926-929}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309109029157}, url = {://WOS:000269398400019}, author = {Kladova, A. V. and Gavel, O. Yu and Mukhopaadhyay, A. and Boer, D. R. and Teixeira, S. and Shnyrov, V. L. and Moura, I. and Moura, J. J. G. and Romao, M. J. and Trincao, J. and Bursakov, S. A.} } @article {7251, title = {Crystallization and crystallographic analysis of the apo form of the orange protein (ORP) from Desulfovibrio gigas}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {65}, year = {2009}, note = {

Times Cited: 2 Romao, Maria/A-4115-2013; Moura, Isabel/D-6339-2013; Moura, Jose/D-6426-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 7 2

}, pages = {730-732}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309109023392}, url = {://WOS:000267530000020}, author = {Najmudin, Shabir and Bonifacio, Cecilia and Duarte, Americo G. and Pualeta, Sofia R. and Moura, Isabel and Moura, Jose J. G. and Romao, Maria J.} } @article {7250, title = {Crystallization and crystallographic analysis of the apo form of the orange protein (ORP) from Desulfovibrio gigas. (vol F65, pg 730, 2009)}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {65}, year = {2009}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Pauleta, Sofia/F-3619-2011; Moura, Isabel/D-6339-2013; Moura, Jose/D-6426-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Pauleta, Sofia/0000-0002-2149-9416; Moura, Isabel/0000-0003-0971-4977; 8 0

}, pages = {856-856}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s174430910902644x}, url = {://WOS:000268517300030}, author = {Najmudin, Shabir and Bonifacio, Cecilia and Duarte, Americo G. and Pauleta, Sofia R. and Moura, Isabel and Moura, Jose J. G. and Romao, Maria J.} } @article {7196, title = {The Effect of the Sixth Sulfur Ligand in the Catalytic Mechanism of Periplasmic Nitrate Reductase}, journal = {Journal of Computational Chemistry}, volume = {30}, number = {15}, year = {2009}, note = {

Times Cited: 14 Romao, Carlos /D-1162-2010; Cerqueira, Nuno M. F. Sousa A./A-5182-2008; Romao, Maria/A-4115-2013; Moura, Isabel/D-6339-2013; Moura, Jose/D-6426-2013; Gonzalez, Pablo/D-8200-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, TCB/M-6190-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Carlos /0000-0001-5061-3743; Cerqueira, Nuno M. F. Sousa A./0000-0003-0342-7424; Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 14

}, pages = {2466-2484}, type = {Journal Article}, abstract = {n/a}, issn = {0192-8651}, doi = {10.1002/jcc.21280}, url = {://WOS:000270869600009}, author = {Cerqueira, N. M. F. S. A. and Gonzalez, P. J. and Brondino, C. D. and Romao, M. J. and Romao, C. C. and Moura, I. and Moura, J. J. G.} } @article {7289, title = {Kinetic, Structural, and EPR Studies Reveal That Aldehyde Oxidoreductase from Desulfovibrio gigas Does Not Need a Sulfido Ligand for Catalysis and Give Evidence for a Direct Mo-C Interaction in a Biological System}, journal = {Journal of the American Chemical Society}, volume = {131}, number = {23}, year = {2009}, note = {

Times Cited: 12 Thapper, Anders/A-6728-2010; Romao, Maria/A-4115-2013; Moura, Isabel/D-6339-2013; Moura, Jose/D-6426-2013; Gonzalez, Pablo/D-8200-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 12

}, pages = {7990-7998}, type = {Journal Article}, abstract = {n/a}, issn = {0002-7863}, doi = {10.1021/ja809448r}, url = {://WOS:000267623100020}, author = {Santos-Silva, Teresa and Ferroni, Felix and Thapper, Anders and Marangon, Jacopo and Gonzalez, Pablo J. and Rizzi, Alberto C. and Moura, Isabel and Moura, Jose J. G. and Romao, Maria J. and Brondino, Carlos D.} } @article {7278, title = {Molybdenum and tungsten enzymes: a crystallographic and mechanistic overview}, journal = {Dalton Transactions}, number = {21}, year = {2009}, note = {

Times Cited: 49 Romao, Maria/A-4115-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 49

}, pages = {4053-4068}, type = {Journal Article}, abstract = {n/a}, issn = {1477-9226}, doi = {10.1039/b821108f}, url = {://WOS:000266212800001}, author = {Romao, Maria Joao} } @article {7219, title = {Preliminary structural characterization of human SOUL, a haem-binding protein}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {65}, year = {2009}, note = {

Times Cited: 2 Freire, Filipe/F-6505-2010; Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; Macedo, Anjos/D-7216-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Freire, Filipe/0000-0002-5310-0668; Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Macedo, Anjos/0000-0002-2613-4838; 2

}, pages = {723-726}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s174430910902291x}, url = {://WOS:000267530000018}, author = {Freire, Filipe and Romao, Maria Joao and Macedo, Anjos L. and Aveiro, Susana S. and Goodfellow, Brian J. and Carvalho, Ana Luisa} } @article {7299, title = {Real-Time Monitoring of Molecular Dynamics of Ethylene Glycol Dimethacrylate Glass Former}, journal = {Journal of Physical Chemistry B}, volume = {113}, number = {43}, year = {2009}, note = {

Times Cited: 11 Salmeron-Sanchez, Manuel/E-4680-2010; Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; Dionisio, Madalena/D-1946-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, CAT/M-4526-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, NSF/N-1636-2013; REQUIMTE, UCIBIO/N-9846-2013; REQUIMTE, LAQV/N-9835-2013 Salmeron-Sanchez, Manuel/0000-0002-8112-2100; Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Dionisio, Madalena/0000-0002-1487-0889; 11

}, pages = {14209-14217}, type = {Journal Article}, abstract = {n/a}, issn = {1520-6106}, doi = {10.1021/jp903212g}, url = {://WOS:000270911100022}, author = {M. T. Viciosa and N. T. Correia and Salmeron Sanchez, M. and Carvalho, A. L. and Romao, M. J. and Gomez Ribelles, J. L. and Dionisio, M.} } @article {7231, title = {Ring-Functionalized Molybdenocene Complexes}, journal = {Organometallics}, volume = {28}, number = {9}, year = {2009}, note = {

Times Cited: 10 Romao, Carlos /D-1162-2010; Romao, Maria/A-4115-2013; Mukhopadhyay, Abhik/D-2119-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Honzicek, Jan/I-6873-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Carlos /0000-0001-5061-3743; Romao, Maria/0000-0002-3004-0543; Mukhopadhyay, Abhik/0000-0002-7755-0401; 10

}, pages = {2871-2879}, type = {Journal Article}, abstract = {n/a}, issn = {0276-7333}, doi = {10.1021/om9001022}, url = {://WOS:000265746100027}, author = {Honzicek, Jan and Mukhopadhyay, Abhik and Santos-Silva, Teresa and Romao, Maria J. and Romao, Carlos C.} } @article {7218, title = {Structural and dynamic characterization of hSOUL, a heme-binding protein}, journal = {Febs Journal}, volume = {276}, year = {2009}, note = {

Times Cited: 0 Freire, Filipe/F-6505-2010; Romao, Maria/A-4115-2013; Macedo, Anjos/D-7216-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Freire, Filipe/0000-0002-5310-0668; Romao, Maria/0000-0002-3004-0543; Macedo, Anjos/0000-0002-2613-4838; Carvalho, Ana Luisa/0000-0002-3824-0240; 34th Congress of the Federation-of-European-Biochemical-Societies Jul 04-09, 2009 Prague, CZECH REPUBLIC Federat European Biochem Soc 0

}, pages = {139-140}, type = {Journal Article}, abstract = {n/a}, issn = {1742-464X}, url = {://WOS:000267069900390}, author = {Freire, F. and Macedo, A. L. and Aveiro, S. S. and Romao, M. J. and Carvalho, A. L. and Goodfellow, B. J.} } @article {7197, title = {Biologically relevant O,S-donor compounds. Synthesis, molybdenum complexation and xanthine oxidase inhibition}, journal = {Dalton Transactions}, number = {13}, year = {2008}, note = {

Times Cited: 9 Chaves, Silvia/H-9865-2012; Santos, M. Amelia /H-9409-2012; Romao, Maria/A-4115-2013; Fresco, Paula/D-4845-2013; Diniz, Carmen/D-6293-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Santos, M. Amelia /0000-0002-4069-9368; Romao, Maria/0000-0002-3004-0543; Fresco, Paula/0000-0001-6705-7798; Diniz, Carmen/0000-0003-4668-9360; 9

}, pages = {1773-1782}, type = {Journal Article}, abstract = {n/a}, issn = {1477-9226}, doi = {10.1039/017172b}, url = {://WOS:000255035500016}, author = {Chaves, Silvia and Gil, Marco and Canario, Sonia and Jelic, Ratomir and Romao, Maria Joao and Trincao, Jose and Herdtweck, Eberhardt and Sousa, Joana and Diniz, Carmen and Fresco, Paula and Santos, M. Amelia} } @article {7300, title = {Ion jelly: a tailor-made conducting material for smart electrochemical devices}, journal = {Chemical Communications}, number = {44}, year = {2008}, note = {

Times Cited: 20 Cabral, Joaquim /G-2052-2010; Barreiros, Susana/A-7892-2012; Lourenco, Nuno/K-5088-2012; Romao, Maria/A-4115-2013; Dionisio, Madalena/D-1946-2013; Mukhopadhyay, Abhik/D-2119-2013; vidinha, Pedro/H-3426-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, CAT/M-4526-2013; REQUIMTE, PHOTO/M-4229-2013; Group, Photochem\&SC/M-4665-2013; REQUIMTE, CENG/M-4555-2013; REQUIMTE, SMB/M-5694-2013; Mateus Afonso, Carlos Alberto/M-7833-2013; Parola, A. Jorge/F-4048-2010; REQUIMTE, NSF/N-1636-2013; REQUIMTE, UCIBIO/N-9846-2013; REQUIMTE, LAQV/N-9835-2013; PTMS, RNEM/C-1589-2014; iMed.ULisboa, iMed.ULisboa/C-6292-2014; iMed.ULisboa, BioOrgChem /B-4209-2014 Cabral, Joaquim /0000-0002-2405-5845; Barreiros, Susana/0000-0002-7268-5892; Lourenco, Nuno/0000-0001-9329-2724; Romao, Maria/0000-0002-3004-0543; Dionisio, Madalena/0000-0002-1487-0889; Mukhopadhyay, Abhik/0000-0002-7755-0401; vidinha, Pedro/0000-0002-3907-4969; Mateus Afonso, Carlos Alberto/0000-0002-7284-5948; Parola, A. Jorge/0000-0002-1333-9076; 20

}, pages = {5842-5844}, type = {Journal Article}, abstract = {n/a}, issn = {1359-7345}, doi = {10.1039/b811647d}, url = {://WOS:000260889800052}, author = {Vidinha, Pedro and Lourenco, Nuno M. T. and Pinheiro, Carlos and Bras, Ana R. and Carvalho, Tania and Santos-Silva, Teresa and Mukhopadhyay, Abhik and Romao, Maria J. and Parola, Jorge and Dionisio, Madalena and Cabral, Joaquim M. S. and Afonso, Carlos A. M. and Barreiros, Susana} } @article {7301, title = {Molecular determinants of ligand specificity in family 11 carbohydrate binding modules - an NMR, X-ray crystallography and computational chemistry approach}, journal = {Febs Journal}, volume = {275}, number = {10}, year = {2008}, note = {

Times Cited: 13 Cabrita, Eurico/E-8053-2011; Bruix, Marta/H-4161-2011; Cerqueira, Nuno M. F. Sousa A./A-5182-2008; Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; Fernandes, Pedro/D-6327-2013; Bras, Natercia/D-5493-2013; Macedo, Anjos/D-7216-2013; Ramos, Maria/D-6183-2013; Viegas, Aldino/A-1889-2011; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, TCB/M-6190-2013; REQUIMTE, UCIBIO/N-9846-2013 Cabrita, Eurico/0000-0002-0720-2751; Cerqueira, Nuno M. F. Sousa A./0000-0003-0342-7424; Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Fernandes, Pedro/0000-0003-2748-4722; Bras, Natercia/0000-0002-3130-9807; Macedo, Anjos/0000-0002-2613-4838; Ramos, Maria/0000-0002-7554-8324; Viegas, Aldino/0000-0003-1733-136X; Prates, Jose/0000-0003-1032-5987; 13

}, pages = {2524-2535}, type = {Journal Article}, abstract = {n/a}, issn = {1742-464X}, doi = {10.1111/j.1742-4658.2008.06401.x}, url = {://WOS:000255285700016}, author = {Viegas, Aldino and Bras, Natercia F. and Cerqueira, Nuno M. F. S. A. and Fernandes, Pedro Alexandrino and Prates, Jose A. M. and Fontes, Carlos M. G. A. and Bruix, Marta and Romao, Maria Joao and Carvalho, Ana Luisa and Ramos, Maria Joao and Macedo, Anjos L. and Cabrita, Eurico J.} } @article {7252, title = {Periplasmic nitrate reductase revisited: a sulfur atom completes the sixth coordination of the catalytic molybdenum}, journal = {Journal of Biological Inorganic Chemistry}, volume = {13}, number = {5}, year = {2008}, note = {

Times Cited: 37 Romao, Carlos /D-1162-2010; Cerqueira, Nuno M. F. Sousa A./A-5182-2008; Romao, Maria/A-4115-2013; Coelho, Catarina /D-2073-2013; Mukhopadhyay, Abhik/D-2119-2013; Moura, Isabel/D-6339-2013; Moura, Jose/D-6426-2013; Gonzalez, Pablo/D-8200-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, TCB/M-6190-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Carlos /0000-0001-5061-3743; Cerqueira, Nuno M. F. Sousa A./0000-0003-0342-7424; Romao, Maria/0000-0002-3004-0543; Coelho, Catarina /0000-0003-1852-5406; Mukhopadhyay, Abhik/0000-0002-7755-0401; Moura, Isabel/0000-0003-0971-4977; 37

}, pages = {737-753}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s00775-008-0359-6}, url = {://WOS:000256320900009}, author = {Najmudin, Shabir and Gonzalez, Pablo J. and Trincao, Jose and Coelho, Catarina and Mukhopadhyay, Abhik and Cerqueira, Nuno M. F. S. A. and Romao, Carlos C. and Moura, Isabel and Moura, Jose J. G. and Brondino, Carlos D. and Romao, Maria J.} } @article {7255, title = {Purification, crystallization and crystallographic analysis of Clostridium thermocellum endo-1,4-beta-D-xylanase 10B in complex with xylohexaose}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {64}, year = {2008}, note = {

Times Cited: 2 Romao, Maria/A-4115-2013; Pinheiro, Benedita/D-2055-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Pinheiro, Benedita/0000-0002-3045-077X; Prates, Jose/0000-0003-1032-5987; 8 2

}, pages = {715-718}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309108019696}, url = {://WOS:000258071000009}, author = {Najmudin, Shabir and Pinheiro, Benedita A. and Romao, Maria J. and Prates, Jose A. M. and Fontes, Carlos M. G. A.} } @article {7224, title = {Purification, crystallization and preliminary X-ray diffraction analysis of adenosine triphosphate sulfurylase (ATPS) from the sulfate-reducing bacterium Desulfovibrio desulfuricans ATCC 27774}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {64}, year = {2008}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Moura, Isabel/D-6339-2013; Moura, Jose/D-6426-2013; Caparica, cqfb_staff/H-2611-2013; REQUIMTE, AL/H-9106-2013; Chaves, Pedro/K-1288-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 7 0

}, pages = {593-595}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309108008816}, url = {://WOS:000257249000005}, author = {Gavel, Olga Yu and Kladova, Anna V. and Bursakov, Sergey A. and Dias, Joao M. and Texeira, Susana and Shnyrov, Valery L. and Moura, Jose J. G. and Moura, Isabel and Romao, Maria J. and Trincao, Jose} } @article {7294, title = {Correlating EPR and X-ray structural analysis of arsenite-inhibited forms of aldehyde oxidoreductase}, journal = {Journal of Biological Inorganic Chemistry}, volume = {12}, number = {3}, year = {2007}, note = {

Times Cited: 6 Thapper, Anders/A-6728-2010; Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 6

}, pages = {353-366}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s00775-006-0191-9}, url = {://WOS:000245457700008}, author = {Thapper, Anders and Boer, D. R. and Brondino, Carlos D. and Moura, Jose J. G. and Romao, Maria J.} } @article {7288, title = {Crystal structure of the 16 heme cytochrome from Desulfovibrio gigas: A glycosylated protein in a sulphate-reducing bacterium}, journal = {Journal of Molecular Biology}, volume = {370}, number = {4}, year = {2007}, note = {

Times Cited: 9 Romao, Maria/A-4115-2013; Lampreia, Jorge/A-9927-2012; REQUIMTE, AL/H-9106-2013; Santos-Silva, Teresa/D-2050-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Lampreia, Jorge/0000-0001-8846-0041; Moura, Isabel/0000-0003-0971-4977; 9

}, pages = {659-673}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1016/j.jmb.2007.04.055}, url = {://WOS:000247904400005}, author = {Santos-Silva, Teresa and Dias, Joao Miguel and Dolla, Alain and Durand, Marie-Claire and Goncalves, Luisa L. and Lampreia, Jorge and Moura, Isabel and Romao, Maria Joao} } @article {7195, title = {Evidence for a dual binding mode of dockerin modules to cohesins}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {104}, number = {9}, year = {2007}, note = {

Times Cited: 56 Davies, Gideon/A-9042-2011; Dias, Fernando/I-4861-2012; Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Davies, Gideon/0000-0002-7343-776X; Dias, Fernando/0000-0001-8109-2063; Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Prates, Jose/0000-0003-1032-5987; 57

}, pages = {3089-3094}, type = {Journal Article}, abstract = {n/a}, issn = {0027-8424}, doi = {10.1073/pnas.0611173104}, url = {://WOS:000244661400015}, author = {Carvalho, Ana Luisa and Dias, Fernando M. V. and Nagy, Tibor and Prates, Jose A. M. and Proctor, Mark R. and Smith, Nicola and Bayer, Edward A. and Davies, Gideon J. and Ferreira, Luis M. A. and Romao, Maria J. and Fontes, Carlos M. G. A. and Gilbert, Harry J.} } @article {7200, title = {Heterodimeric nitrate reductase (NapAB) from Cupriavidus necator H16: purification, crystallization and preliminary X-ray analysis}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {63}, year = {2007}, note = {

Times Cited: 8 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; Coelho, Catarina /D-2073-2013; Moura, Jose/D-6426-2013; Gonzalez, Pablo/D-8200-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Coelho, Catarina /0000-0003-1852-5406; Moura, Isabel/0000-0003-0971-4977; 6 8

}, pages = {516-519}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309107022129}, url = {://WOS:000248045100015}, author = {Coelho, Catarina and Gonzalez, Pablo J. and Trincao, Jose and Carvalho, Ana L. and Najmudin, Shabir and Hettman, Thomas and Dieckman, Stephan and Moura, Jose J. G. and Moura, Isabel and Romao, Maria J.} } @article {7221, title = {The human carnitine acylcarnitine translocase (hCACT): Strategies for its heterologous expression, purification and crystallization}, journal = {Journal of Inherited Metabolic Disease}, volume = {30}, year = {2007}, note = {

Times Cited: 0 LNEG, Producao Cientifica/D-2212-2012; Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013; iMed.ULisboa, MetGen /B-5293-2014; iMed.ULisboa, iMed.ULisboa/C-6292-2014; Soveral, Graca/L-1550-2013; Leandro, Paula/A-8323-2014 Romao, Maria/0000-0002-3004-0543; Carvalho, Ana Luisa/0000-0002-3824-0240; Soveral, Graca/0000-0001-8487-110X; Leandro, Paula/0000-0002-2946-9342 1 0

}, pages = {53-53}, type = {Journal Article}, abstract = {n/a}, issn = {0141-8955}, url = {://WOS:000248860000209}, author = {Fv, Ventura and Violante, S. and Gomes, C. and Carvalho, A. L. and Romao, M. J. and Gaspar, M. M. and Cruz, M. E. M. and Soveral, G. and Wanders, R. J. and Leandro, P. and de Almeida, Tavares V.} } @article {7187, title = {Structural and electron paramagnetic resonance (EPR) studies of mononuclear molybdenum enzymes from sulfate-reducing bacteria (vol 39, pg 793, 2006)}, journal = {Accounts of Chemical Research}, volume = {40}, number = {3}, year = {2007}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 0

}, pages = {231-231}, type = {Journal Article}, abstract = {n/a}, issn = {0001-4842}, doi = {10.1021/ar600049k}, url = {://WOS:000245013100008}, author = {Brondino, Carlos D. and Rivas, Maria G. and Romao, Maria J. and Moura, Jose J. G. and Moura, Isabel} } @article {7287, title = {The first crystal structure of class III superoxide reductase from Treponema pallidum}, journal = {Journal of Biological Inorganic Chemistry}, volume = {11}, number = {5}, year = {2006}, note = {

Times Cited: 24 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, AL/H-9106-2013; Santos-Silva, Teresa/D-2050-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 24

}, pages = {548-558}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s00775-006-0104-y}, url = {://WOS:000238499000002}, author = {Santos-Silva, T. and Trincao, J. and Carvalho, A. L. and Bonifacio, C. and Auchere, F. and Raleiras, P. and Moura, I. and Moura, J. J. G. and Romao, M. J.} } @article {7263, title = {Formate-reduced E-coli formate dehydrogenase H: the reinterpretation of the crystal structure suggests a new reaction mechanism}, journal = {Journal of Biological Inorganic Chemistry}, volume = {11}, number = {7}, year = {2006}, note = {

Times Cited: 37 Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 37

}, pages = {849-854}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s00775-006-0129-2}, url = {://WOS:000240283800005}, author = {Raaijmakers, Hans C. A. and Romao, Maria Joao} } @article {7185, title = {Molybdenum and tungsten enzymes: the xanthine oxidase family}, journal = {Current Opinion in Chemical Biology}, volume = {10}, number = {2}, year = {2006}, note = {

Times Cited: 41 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 41

}, pages = {109-114}, type = {Journal Article}, abstract = {n/a}, issn = {1367-5931}, doi = {10.1016/j.cbpa.2006.01.034}, url = {://WOS:000237000700004}, author = {Brondino, C. D. and Romao, M. J. and Moura, I. and Moura, J. J. G.} } @article {7297, title = {Purification, crystallization and preliminary X-ray diffraction analysis of the glyoxalase II from Leishmania infantum}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {62}, year = {2006}, note = {

Times Cited: 2 Lima, Marta/C-7042-2009; Ferreira, Antonio/E-6488-2012; Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; Tomas, Ana/J-9919-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013; PTMS, RNEM/C-1589-2014 Ferreira, Antonio/0000-0002-9625-8115; Romao, Maria/0000-0002-3004-0543; 8 2

}, pages = {805-807}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309106027539}, url = {://WOS:000239357900025}, author = {Trincao, Jose and Silva, Marta Sousa and Barata, Lidia and Bonifacio, Cecilia and Carvalho, Sandra and Tomas, Ana Maria and Ferreira, Antonio E. N. and Cordeiro, Carlos and Freire, Ana Ponces and Romao, Maria Joao} } @inbook {7277, title = {The role of molybdenum in biology}, booktitle = {Metal Ions in Biology and Medicine, Vol 9}, series = {Metal Ions in Biology and Medicine}, volume = {9}, year = {2006}, note = {

Times Cited: 1 Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 9th International Symposium on Metal Ions in Biology and Medicine May 21-24, 2006 Lisbon, PORTUGAL 1

}, pages = {507-510}, type = {Book Section}, abstract = {n/a}, isbn = {1257-2535 2-7420-0629-X}, url = {://WOS:000242710200090}, author = {Romao, Maria Joao}, editor = {Alpoim, M.C. and Morais, P.V.} } @article {7186, title = {Structural and electron paramagnetic resonance (EPR) studies of mononuclear molybdenum enzymes from sulfate-reducing bacteria}, journal = {Accounts of Chemical Research}, volume = {39}, number = {10}, year = {2006}, note = {

Times Cited: 28 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; Rivas, Maria/D-7883-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 28

}, pages = {788-796}, type = {Journal Article}, abstract = {n/a}, issn = {0001-4842}, doi = {10.1021/ar050104k}, url = {://WOS:000241325500012}, author = {Brondino, Carlos D. and Rivas, Maria G. and Romao, Maria J. and Moura, Jose J. G. and Moura, Isabel} } @article {7249, title = {Xyloglucan is recognized by carbohydrate-binding modules that interact with beta-glucan chains}, journal = {Journal of Biological Chemistry}, volume = {281}, number = {13}, year = {2006}, note = {

Times Cited: 42 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; Prates, Jose/K-9934-2013; Correia, Marcia/D-2077-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Prates, Jose/0000-0003-1032-5987; Correia, Marcia/0000-0003-0636-8095; 42

}, pages = {8815-8828}, type = {Journal Article}, abstract = {n/a}, issn = {0021-9258}, doi = {10.1074/jbc.M510559200}, url = {://WOS:000236247100061}, author = {Najmudin, S. and Guerreiro, Cipd and Carvalho, A. L. and Prates, J. A. M. and Correia, M. A. S. and Alves, V. D. and Ferreira, L. M. A. and Romao, M. J. and Gilbert, H. J. and Bolam, D. N. and Fontes, Cmga} } @article {7193, title = {Insights into the structural determinants of cohesin dockerin specificity revealed by the crystal structure of the type II cohesin from Clostridium thermocellum SdbA}, journal = {Journal of Molecular Biology}, volume = {349}, number = {5}, year = {2005}, note = {

Times Cited: 25 Davies, Gideon/A-9042-2011; Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; Pires, Virginia/J-4799-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Davies, Gideon/0000-0002-7343-776X; Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Pires, Virginia/0000-0003-1307-3797; Prates, Jose/0000-0003-1032-5987; 25

}, pages = {909-915}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1016/j.jmb.2005.04.037}, url = {://WOS:000229805300001}, author = {Carvalho, A. L. and Pires, V. M. R. and Gloster, T. M. and Turkenburg, J. P. and Prates, J. A. M. and Ferreira, L. M. A. and Romao, M. J. and Davies, G. J. and Fontes, Cmga and Gilbert, H. J.} } @article {7178, title = {On the purification and preliminary crystallographic analysis of isoquinoline 1-oxidoreductase from Brevundimonas diminuta 7}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {61}, year = {2005}, note = {

Times Cited: 3 Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 1 3

}, pages = {137-140}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s1744309104032105}, url = {://WOS:000232157000040}, author = {Boer, D. R. and Muller, A. and Fetzner, S. and Lowe, D. J. and Romao, M. J.} } @article {7286, title = {Superoxide reductase from the syphilis spirochete Treponema pallidum: crystallization and structure determination using soft X-rays}, journal = {Acta Crystallographica Section F-Structural Biology and Crystallization Communications}, volume = {61}, year = {2005}, note = {

Times Cited: 4 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, AL/H-9106-2013; Santos-Silva, Teresa/D-2050-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 11 4

}, pages = {967-970}, type = {Journal Article}, abstract = {n/a}, issn = {1744-3091}, doi = {10.1107/s174430910502885x}, url = {://WOS:000232890200003}, author = {Santos-Silva, T. and Trincao, J. and Carvalho, A. L. and Bonifacio, C. and Auchere, F. and Moura, I. and Moura, J. J. G. and Romao, M. J.} } @article {7284, title = {Crystallization and preliminary X-ray diffraction analysis of the 16-haem cytochrome of Desulfovibrio gigas}, journal = {Acta Crystallographica Section D-Biological Crystallography}, volume = {60}, year = {2004}, note = {

Times Cited: 2 Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; Santos-Silva, Teresa/D-2050-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 5 2

}, pages = {968-970}, type = {Journal Article}, abstract = {n/a}, issn = {0907-4449}, doi = {10.1107/s0907444904005621}, url = {://WOS:000220951000032}, author = {Santos-Silva, T. and Diasa, J. M. and Bourenkov, G. and Bartunik, H. and Moura, I. and Romao, M. J.} } @article {7213, title = {Direct electrochemistry of the Desulfovibrio gigas aldehyde oxidoreductase}, journal = {European Journal of Biochemistry}, volume = {271}, number = {7}, year = {2004}, note = {

Times Cited: 10 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, AL/H-9106-2013; dos Santos, Margarida/H-7897-2012; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; dos Santos, Margarida/0000-0001-7531-757X; Moura, Isabel/0000-0003-0971-4977; 10

}, pages = {1329-1338}, type = {Journal Article}, abstract = {n/a}, issn = {0014-2956}, doi = {10.1111/j.1432-1033.2004.04041.x}, url = {://WOS:000220355100011}, author = {dos Santos, M. M. C. and Sousa, P. M. P. and Goncalves, M. L. S. and Romao, M. J. and Moura, I. and Moura, J. J. G.} } @article {7192, title = {The family 11 carbohydrate-binding module of Clostridium thermocellum Lic26A-Cel5E accommodates beta-1,4- and beta-1,3-1,4-mixed linked glucans at a single binding site}, journal = {Journal of Biological Chemistry}, volume = {279}, number = {33}, year = {2004}, note = {

Times Cited: 51 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; Pires, Virginia/J-4799-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Pires, Virginia/0000-0003-1307-3797; Prates, Jose/0000-0003-1032-5987; 57

}, pages = {34785-34793}, type = {Journal Article}, abstract = {n/a}, issn = {0021-9258}, doi = {10.1074/jbc.M405867200}, url = {://WOS:000223134800086}, author = {Carvalho, A. L. and Goyal, A. and Prates, J. A. M. and Bolam, D. N. and Gilbert, H. J. and Pires, V. M. R. and Ferreira, L. M. A. and Planas, A. and Romao, M. J. and Fontes, Cmga} } @article {7246, title = {Mo and W bis-MGD enzymes: nitrate reductases and formate dehydrogenases}, journal = {Journal of Biological Inorganic Chemistry}, volume = {9}, number = {7}, year = {2004}, note = {

Times Cited: 70 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 74

}, pages = {791-799}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s00775-004-0573-9}, url = {://WOS:000224751400001}, author = {Moura, J. J. G. and Brondino, C. D. and Trincao, J. and Romao, M. J.} } @article {7229, title = {Mutagenesis study on amino acids around the molybdenum centre of the periplasmic nitrate reductase from Ralstonia eutropha}, journal = {Biochemical and Biophysical Research Communications}, volume = {320}, number = {4}, year = {2004}, note = {

Times Cited: 11 Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; Santos-Silva, Teresa/D-2050-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 12

}, pages = {1211-1219}, type = {Journal Article}, abstract = {n/a}, issn = {0006-291X}, doi = {10.1016/j.bbrc.2004.06.086}, url = {://WOS:000222923400025}, author = {Hettmann, T. and Siddiqui, R. A. and Frey, C. and Santos-Silva, T. and Romao, M. J. and Diekmann, S.} } @article {7206, title = {Structural basis for the mechanism of Ca2+ activation of the di-heme cytochrome c peroxidase from Pseudomonas nautica 617}, journal = {Structure}, volume = {12}, number = {6}, year = {2004}, note = {

Times Cited: 38 Romao, Maria/A-4115-2013; Pereira, Alice/D-7779-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 38

}, pages = {961-973}, type = {Journal Article}, abstract = {n/a}, issn = {0969-2126}, doi = {10.1016/j.str.2004.03.025}, url = {://WOS:000222155100009}, author = {Dias, J. M. and Alves, T. and Bonifacio, C. and Pereira, A. S. and Trincao, J. and Bourgeois, D. and Moura, I. and Romao, M. J.} } @article {7179, title = {X-ray crystal structure and EPR spectra of "arsenite-inhibited" Desulfovibrio gigas aldehyde dehydrogenase: A member of the xanthine oxidase family}, journal = {Journal of the American Chemical Society}, volume = {126}, number = {28}, year = {2004}, note = {

Times Cited: 14 Thapper, Anders/A-6728-2010; Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 14

}, pages = {8614-8615}, type = {Journal Article}, abstract = {n/a}, issn = {0002-7863}, doi = {10.1021/ja0490222}, url = {://WOS:000222704700012}, author = {Boer, D. R. and Thapper, A. and Brondino, C. D. and Romao, M. J. and Moura, J. J. G.} } @article {7190, title = {Cellulosome assembly revealed by the crystal structure of the cohesin-dockerin complex}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {100}, number = {24}, year = {2003}, note = {

Times Cited: 130 Davies, Gideon/A-9042-2011; Dias, Fernando/I-4861-2012; Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; Prates, Jose/K-9934-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Davies, Gideon/0000-0002-7343-776X; Dias, Fernando/0000-0001-8109-2063; Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; Prates, Jose/0000-0003-1032-5987; 131

}, pages = {13809-13814}, type = {Journal Article}, abstract = {n/a}, issn = {0027-8424}, doi = {10.1073/pnas.1936124100}, url = {://WOS:000186803800015}, author = {Carvalho, A. L. and Dias, F. M. V. and Prates, J. A. M. and Nagy, T. and Gilbert, H. J. and Davies, G. J. and Ferreira, L. M. A. and Romao, M. J. and Fontes, Cmga} } @article {7180, title = {Crystallization and preliminary X-ray diffraction analysis of the di-haem cytochrome c peroxidase from Pseudomonas stutzeri}, journal = {Acta Crystallographica Section D-Biological Crystallography}, volume = {59}, year = {2003}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; Timoteo, Cristina/D-7275-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 2 0

}, pages = {345-347}, type = {Journal Article}, abstract = {n/a}, issn = {0907-4449}, doi = {10.1107/s0907444902020760}, url = {://WOS:000180641900018}, author = {Bonifacio, C. and Cunha, C. A. and Muller, A. and Timoteo, C. G. and Dias, J. M. and Moura, I. and Romao, M. J.} } @article {7204, title = {Cytochrome c nitrite reductase from Desulfovibrio desulfuricans ATCC 27774 - The relevance of the two calcium sites in the structure of the catalytic subunit (NrfA)}, journal = {Journal of Biological Chemistry}, volume = {278}, number = {19}, year = {2003}, note = {

Times Cited: 47 Costa, Cristina/A-9966-2012; Romao, Maria/A-4115-2013; Lampreia, Jorge/A-9927-2012; Moura, Jose/D-6426-2013; Almeida, Maria Gabriela/D-8265-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Costa, Cristina/0000-0002-8611-9023; Romao, Maria/0000-0002-3004-0543; Lampreia, Jorge/0000-0001-8846-0041; Almeida, Maria Gabriela/0000-0002-4508-7379; Moura, Isabel/0000-0003-0971-4977; 47

}, pages = {17455-17465}, type = {Journal Article}, abstract = {n/a}, issn = {0021-9258}, doi = {10.1074/jbc.M211777200}, url = {://WOS:000182818600133}, author = {Cunha, C. A. and Macieira, S. and Dias, J. M. and Almeida, G. and Goncalves, L. L. and Costa, C. and Lampreia, J. and Huber, R. and Moura, J. J. G. and Moura, I. and Romao, M. J.} } @article {7173, title = {The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774 - Re-evaluation of the spectroscopic data and redox properties}, journal = {European Journal of Biochemistry}, volume = {270}, number = {19}, year = {2003}, note = {

Times Cited: 32 Costa, Cristina/A-9966-2012; Romao, Maria/A-4115-2013; Lampreia, Jorge/A-9927-2012; Moura, Jose/D-6426-2013; Almeida, Maria Gabriela/D-8265-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Costa, Cristina/0000-0002-8611-9023; Romao, Maria/0000-0002-3004-0543; Lampreia, Jorge/0000-0001-8846-0041; Almeida, Maria Gabriela/0000-0002-4508-7379; Moura, Isabel/0000-0003-0971-4977; 32

}, pages = {3904-3915}, type = {Journal Article}, abstract = {n/a}, issn = {0014-2956}, doi = {10.1046/j.1432-1033.2003.03772.x}, url = {://WOS:000185370100005}, author = {Almeida, M. G. and Macieira, S. and Goncalves, L. L. and Huber, R. and Cunha, C. A. and Romao, M. J. and Costa, C. and Lampreia, J. and Moura, J. J. G. and Moura, I.} } @article {7222, title = {Mammalian molybdo-flavoenzymes, an expanding family of proteins: structure, genetics, regulation, function and pathophysiology}, journal = {Biochemical Journal}, volume = {372}, year = {2003}, note = {

Times Cited: 110 Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 1 116

}, pages = {15-32}, type = {Journal Article}, abstract = {n/a}, issn = {0264-6021}, doi = {10.1042/bj20030121}, url = {://WOS:000183070800002}, author = {Garattini, E. and Mendel, R. and Romao, M. J. and Wright, R. and Terao, M.} } @article {7245, title = {Molecular aspects of denitrification/nitrate dissimilation}, journal = {Journal of Inorganic Biochemistry}, volume = {96}, number = {1}, year = {2003}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; Almeida, Maria Gabriela/D-8265-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Almeida, Maria Gabriela/0000-0002-4508-7379; Moura, Isabel/0000-0003-0971-4977; 11th International Conference on Biological Inorganic Chemistry Jul 19-23, 2003 Caims, australia 0

}, pages = {195-195}, type = {Journal Article}, abstract = {n/a}, issn = {0162-0134}, doi = {10.1016/s0162-0134(03)80717-1}, url = {://WOS:000184009800306}, author = {Moura, I. and Cabrito, I. and Almeida, G. and Cunha, C. and Romao, M. J. and Moura, J. J. G.} } @article {7230, title = {Mutagenesis study on the role of a lysine residue highly conserved in formate dehydrogenases and periplasmic nitrate reductases}, journal = {Biochemical and Biophysical Research Communications}, volume = {310}, number = {1}, year = {2003}, note = {

Times Cited: 12 Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 12

}, pages = {40-47}, type = {Journal Article}, abstract = {n/a}, issn = {0006-291X}, doi = {10.1016/j.bbrc.2003.08.114}, url = {://WOS:000185835500007}, author = {Hettmann, T. and Siddiqui, R. A. and van Langen, J. and Frey, C. and Romao, M. J. and Diekmann, S.} } @article {7194, title = {Crystal structure of a prostate kallikrein isolated from stallion seminal plasma: A homologue of human PSA}, journal = {Journal of Molecular Biology}, volume = {322}, number = {2}, year = {2002}, note = {

Times Cited: 51 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; 52

}, pages = {325-337}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1016/s0022-2836(02)00705-2}, url = {://WOS:000178230500007}, author = {Carvalho, A. L. and Sanz, L. and Barettino, D. and Romero, A. and Calvete, J. J. and Romao, M. J.} } @article {7207, title = {Crystallization and preliminary X-ray diffraction analysis of two pH-dependent forms of a di-haem cytochrome c peroxidase from Pseudomonas nautica}, journal = {Acta Crystallographica Section D-Biological Crystallography}, volume = {58}, year = {2002}, note = {

Times Cited: 4 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 4

}, pages = {697-699}, type = {Journal Article}, abstract = {n/a}, issn = {0907-4449}, doi = {10.1107/s0907444902002354}, url = {://WOS:000174727700022}, author = {Dias, J. M. and Bonifacio, C. and Alves, T. and Moura, J. J. G. and Moura, I. and Romao, M. J.} } @article {7261, title = {Gene sequence and the 1.8 angstrom crystal structure of the tungsten-containing formate dehydrogenase from Desulfolvibrio gigas}, journal = {Structure}, volume = {10}, number = {9}, year = {2002}, note = {

Times Cited: 66 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 72

}, pages = {1261-1272}, type = {Journal Article}, abstract = {n/a}, issn = {0969-2126}, doi = {10.1016/s0969-2126(02)00826-2}, url = {://WOS:000178030800014}, author = {Raaijmakers, H. and Macieira, S. and Dias, J. M. and Teixeira, S. and Bursakov, S. and Huber, R. and Moura, J. J. G. and Moura, I. and Romao, M. J.} } @article {7270, title = {Molybdenum enzymes in reactions involving aldehydes and acids}, journal = {Molybdenum and Tungsten: Their Roles in Biological Processes}, volume = {39}, year = {2002}, note = {

Times Cited: 8 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 8

}, pages = {539-570}, type = {Journal Article}, abstract = {n/a}, issn = {0161-5149}, url = {://WOS:000175393800015}, author = {Romao, M. J. and Cunha, C. A. and Brondino, C. D. and Moura, J. J. G.} } @article {7191, title = {Purification, crystallization and identification by X-ray analysis of a prostate kallikrein from horse seminal plasma}, journal = {Acta Crystallographica Section D-Biological Crystallography}, volume = {57}, year = {2001}, note = {

Times Cited: 2 Romao, Maria/A-4115-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Carvalho, Ana Luisa/0000-0002-3824-0240; 8 2

}, pages = {1180-1183}, type = {Journal Article}, abstract = {n/a}, issn = {0907-4449}, doi = {10.1107/s0907444901009805}, url = {://WOS:000170041200025}, author = {Carvalho, A. L. and Dias, J. M. and Sanz, L. and Romero, A. and Calvete, J. J. and Romao, M. J.} } @article {7226, title = {Structural studies on Desulfovibrio desulfuricans ATCC 27774 multiheme nitrite reductase - characterization of the subunits}, journal = {Journal of Inorganic Biochemistry}, volume = {86}, number = {1}, year = {2001}, note = {

Times Cited: 0 Costa, Cristina/A-9966-2012; Romao, Maria/A-4115-2013; Lampreia, Jorge/A-9927-2012; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013; Almeida, Maria Gabriela/D-8265-2013 Costa, Cristina/0000-0002-8611-9023; Romao, Maria/0000-0002-3004-0543; Lampreia, Jorge/0000-0001-8846-0041; Moura, Isabel/0000-0003-0971-4977; Almeida, Maria Gabriela/0000-0002-4508-7379 0

}, pages = {316-316}, type = {Journal Article}, abstract = {n/a}, issn = {0162-0134}, url = {://WOS:000170467300603}, author = {Goncalves, L. M. L. and Cunha, C. and Almeida, G. and Macieira, S. and Costa, C. and Lampreia, J. and Romao, M. J. and Moura, J. J. G. and Moura, I.} } @article {7264, title = {Structure refinement of the aldehyde oxidoreductase from Desulfovibrio gigas (MOP) at 1.28 angstrom}, journal = {Journal of Biological Inorganic Chemistry}, volume = {6}, number = {8}, year = {2001}, note = {

Times Cited: 52 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 54

}, pages = {791-800}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, url = {://WOS:000172413300005}, author = {Rebelo, J. M. and Dias, J. M. and Huber, R. and Moura, J. J. G. and Romao, M. J.} } @article {7262, title = {Tungsten-containing formats dehydrogenase from Desulfovibrio gigas: metal identification and preliminary structural data by multi-wavelength crystallography}, journal = {Journal of Biological Inorganic Chemistry}, volume = {6}, number = {4}, year = {2001}, note = {

Times Cited: 25 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 29

}, pages = {398-404}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s007750100215}, url = {://WOS:000168528400007}, author = {Raaijmakers, H. and Teixeira, S. and Dias, J. M. and Almendra, M. J. and Brondino, C. D. and Moura, I. and Moura, J. J. G. and Romao, M. J.} } @article {7214, title = {Biochemical/spectroscopic characterization and preliminary X-ray analysis of a new aldehyde oxidoreductase isolated from Desulfovibrio desulfuricans ATCC 27774}, journal = {Biochemical and Biophysical Research Communications}, volume = {268}, number = {3}, year = {2000}, note = {

Times Cited: 8 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 8

}, pages = {745-749}, type = {Journal Article}, abstract = {n/a}, issn = {0006-291X}, doi = {10.1006/bbrc.2000.2135}, url = {://WOS:000085640000016}, author = {Duarte, R. O. and Archer, M. and Dias, J. M. and Bursakov, S. and Huber, R. and Moura, I. and Romao, M. J. and Moura, J. J. G.} } @article {7210, title = {Crystallization and preliminary X-ray analysis of a membrane-bound nitrite reductase from Desulfovibrio desulfuricans ATCC 27774}, journal = {Acta Crystallographica Section D-Biological Crystallography}, volume = {56}, year = {2000}, note = {

Times Cited: 6 Costa, Cristina/A-9966-2012; Romao, Maria/A-4115-2013; Lampreia, Jorge/A-9927-2012; Almeida, Maria Gabriela/D-8265-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Costa, Cristina/0000-0002-8611-9023; Romao, Maria/0000-0002-3004-0543; Lampreia, Jorge/0000-0001-8846-0041; Almeida, Maria Gabriela/0000-0002-4508-7379; Moura, Isabel/0000-0003-0971-4977; 2 6

}, pages = {215-217}, type = {Journal Article}, abstract = {n/a}, issn = {0907-4449}, doi = {10.1107/s0907444999016066}, url = {://WOS:000085557600020}, author = {Dias, J. M. and Cunha, C. A. and Teixeira, S. and Almeida, G. and Costa, C. and Lampreia, J. and Moura, J. J. G. and Moura, I. and Romao, M. J.} } @article {7265, title = {Gene sequence and crystal structure of the aldehyde oxidoreductase from Desulfovibrio desulfuricans ATCC 27774}, journal = {Journal of Molecular Biology}, volume = {297}, number = {1}, year = {2000}, note = {

Times Cited: 46 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 46

}, pages = {135-146}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1006/jmbi.2000.3552}, url = {://WOS:000085976500011}, author = {Rebelo, J. and Macieira, S. and Dias, J. M. and Huber, R. and Ascenso, C. S. and Rusnak, F. and Moura, J. J. G. and Moura, I. and Romao, M. J.} } @article {7282, title = {Ionic strength dependence of the non-physiological electron transfer between flavodoxin and cytochrome c(553) from D-vulgaris}, journal = {Journal of Biological Inorganic Chemistry}, volume = {5}, number = {6}, year = {2000}, note = {

Times Cited: 8 GILARDI, GIANFRANCO/B-4523-2010; Soares, Claudio/E-2675-2012; Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 8

}, pages = {730-737}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s007750000162}, url = {://WOS:000165432900007}, author = {Sadeghi, S. J. and Valetti, F. and Cunha, C. A. and Romao, M. J. and Soares, C.M. and Gilardi, G.} } @article {7188, title = {Cross immunological reactions and spectroscopy study within nitrate reductase and other mononuclear Mo containing enzymes of the sulfate reducing bacteria}, journal = {Journal of Inorganic Biochemistry}, volume = {74}, number = {1-4}, year = {1999}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 0

}, pages = {86-86}, type = {Journal Article}, abstract = {n/a}, issn = {0162-0134}, url = {://WOS:000081157700161}, author = {Bursakov, S. A. and Brondino, C. and Dias, J. M. and Carneiro, C. and Caldeira, J. and Duarte, R. O. and Romao, M. J. and Moura, I. and Moura, J. J. G.} } @article {7212, title = {Crystal structure of the first dissimilatory nitrate reductase at 1.9 angstrom solved by MAD methods}, journal = {Structure with Folding \& Design}, volume = {7}, number = {1}, year = {1999}, note = {

Times Cited: 222 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; Szaleniec, Maciej/A-1198-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 226

}, pages = {65-79}, type = {Journal Article}, abstract = {n/a}, issn = {0969-2126}, doi = {10.1016/s0969-2126(99)80010-0}, url = {://WOS:000079349700010}, author = {Dias, J. M. and Than, M. E. and Humm, A. and Huber, R. and Bourenkov, G. P. and Bartunik, H. D. and Bursakov, S. and Calvete, J. and Caldeira, J. and Carneiro, C. and Moura, J. J. G. and Moura, I. and Romao, M. J.} } @article {7208, title = {Crystallization and preliminary X-ray analysis of a nitrate reductase from Desulfovibrio desulfuricans ATCC 27774}, journal = {Acta Crystallographica Section D-Biological Crystallography}, volume = {55}, year = {1999}, note = {

Times Cited: 4 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 4

}, pages = {877-879}, type = {Journal Article}, abstract = {n/a}, issn = {0907-4449}, doi = {10.1107/s0907444998014735}, url = {://WOS:000079855200022}, author = {Dias, J. M. and Bursakov, S. and Carneiro, C. and Moura, J. J. G. and Moura, I. and Romao, M. J.} } @article {7211, title = {Crystallographic studies of a dissimilatory nitrate reductase and mechanistic implications}, journal = {Journal of Inorganic Biochemistry}, volume = {74}, number = {1-4}, year = {1999}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 0

}, pages = {113-113}, type = {Journal Article}, abstract = {n/a}, issn = {0162-0134}, url = {://WOS:000081157700214}, author = {Dias, J. M. and Than, M. E. and Huber, R. and Bourenkov, G. P. and Bartunik, H. D. and Bursakov, S. and Moura, J. J. G. and Moura, I. and Romao, M. J.} } @article {7293, title = {Crystallographic studies on a tungsten-containning formate dehydrogenase from Desulfovibrio gigas}, journal = {Journal of Inorganic Biochemistry}, volume = {74}, number = {1-4}, year = {1999}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Carvalho, Ana Luisa/0000-0002-3824-0240; Moura, Isabel/0000-0003-0971-4977; 0

}, pages = {89-89}, type = {Journal Article}, abstract = {n/a}, issn = {0162-0134}, url = {://WOS:000081157700167}, author = {Teixeira, S. and Dias, J. M. and Carvalho, A. L. and Bourenkov, G. and Bartunik, H. and Almendra, M. J. and Moura, I. and Moura, J. J. G. and Romao, M. J.} } @article {7205, title = {Effects of protein-protein interactions on electron transfer: docking and electron transfer calculations for complexes between flavodoxin and c-type cytochromes}, journal = {Journal of Biological Inorganic Chemistry}, volume = {4}, number = {3}, year = {1999}, note = {

Times Cited: 18 GILARDI, GIANFRANCO/B-4523-2010; Soares, Claudio/E-2675-2012; Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 19

}, pages = {360-374}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, url = {://WOS:000081613300014}, author = {Cunha, C. A. and Romao, M. J. and Sadeghi, S. J. and Valetti, F. and Gilardi, G. and Soares, C.M.} } @article {7269, title = {Preliminary crystallographic studies of xanthine oxidase purified from rat liver}, journal = {Journal of Inorganic Biochemistry}, volume = {74}, number = {1-4}, year = {1999}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Carvalho, Ana Luisa/0000-0002-3824-0240; 0

}, pages = {281-281}, type = {Journal Article}, abstract = {n/a}, issn = {0162-0134}, url = {://WOS:000081157700522}, author = {Romao, M. J. and Carvalho, A. L. and Dias, J. M. and Teixeira, S. and Bourenkov, G. and Bartunik, H. and Huber, R. and Maia, L. and Mira, L.} } @article {7175, title = {Structural studies by X-ray diffraction on metal substituted desulforedoxin, a rubredoxin-type protein}, journal = {Protein Science}, volume = {8}, number = {7}, year = {1999}, note = {

Times Cited: 12 Romao, Maria/A-4115-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Carvalho, Ana Luisa/0000-0002-3824-0240; Moura, Isabel/0000-0003-0971-4977; 13

}, pages = {1536-1545}, type = {Journal Article}, abstract = {n/a}, issn = {0961-8368}, url = {://WOS:000081271700019}, author = {Archer, M. and Carvalho, A. L. and Teixeira, S. and Moura, I. and Moura, J. J. G. and Rusnak, F. and Romao, M. J.} } @article {7225, title = {Altered specificity mutations define residues essential for substrate positioning in xanthine dehydrogenase}, journal = {Journal of Molecular Biology}, volume = {278}, number = {2}, year = {1998}, note = {

Times Cited: 20 Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 22

}, pages = {431-438}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1006/jmbi.1998.1707}, url = {://WOS:000073696500012}, author = {Glatigny, A. and Hof, P. and Romao, M. J. and Huber, R. and Scazzocchio, C.} } @article {7271, title = {Structure and function of the xanthine-oxidase family of molybdenum enzymes}, journal = {Metal Sites in Proteins and Models}, volume = {90}, year = {1998}, note = {

Times Cited: 32 Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 32

}, pages = {69-95}, type = {Journal Article}, abstract = {n/a}, issn = {0081-5993}, url = {://WOS:000073873900003}, author = {Romao, M. J. and Huber, R.} } @article {7304, title = {Substrate oxidation in the active site of xanthine oxidase and related enzymes. A model density functional study}, journal = {Inorganic Chemistry}, volume = {37}, number = {2}, year = {1998}, note = {

Times Cited: 35 Roesch, Notker/C-1182-2010; Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 35

}, pages = {176-180}, type = {Journal Article}, abstract = {n/a}, issn = {0020-1669}, doi = {10.1021/ic9707570}, url = {://WOS:000071713300005}, author = {Voityuk, A. A. and Albert, K. and Romao, M. J. and Huber, R. and Rosch, N.} } @article {7298, title = {The 2.4 angstrom resolution crystal structure of boar seminal plasma PSP-I/PSP-II: a zona pellucida-binding glycoprotein heterodimer of the spermadhesin family built by a CUB domain architecture}, journal = {Journal of Molecular Biology}, volume = {274}, number = {4}, year = {1997}, note = {

Times Cited: 65 Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 65

}, pages = {635-649}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1006/jmbi.1997.1424}, url = {://WOS:000071252400017}, author = {Varela, P. F. and Romero, A. and Sanz, L. and Romao, M. J. and Topfer-Petersen, E. and Calvete, J. J.} } @article {7272, title = {Crystal structure and mechanism of action of the xanthine oxidase-related aldehyde oxidoreductase from Desulfovibrio gigas}, journal = {Biochemical Society Transactions}, volume = {25}, number = {3}, year = {1997}, note = {

Times Cited: 6 Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Symposium on Xanthine Oxidase - Enzymology and Pathophysiology, at the 661st Meeting of the Biochemical-Society Apr 10-11, 1997 Bath, england Biochem Soc; Soc Free Rad Res 6

}, pages = {755-757}, type = {Journal Article}, abstract = {n/a}, issn = {0300-5127}, url = {://WOS:A1997XV20900003}, author = {Romao, M. J. and Hubert, R.} } @article {7274, title = {Crystal structure of acidic seminal fluid protein (aSFP) at 1.9 angstrom resolution: a bovine polypeptide of the spermadhesin family}, journal = {Journal of Molecular Biology}, volume = {274}, number = {4}, year = {1997}, note = {

Times Cited: 34 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; 34

}, pages = {650-660}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1006/jmbi.1997.1423}, url = {://WOS:000071252400018}, author = {Romao, M. J. and Kolln, I. and Dias, J. M. and Carvalho, A. L. and Romero, A. and Varela, P. F. and Sanz, L. and Topfer-Petersen, E. and Calvete, J. J.} } @article {7174, title = {Crystal structure of cytochrome c{\textquoteright} from Rhodocyclus gelatinosus and comparison with other cytochromes c{\textquoteright}}, journal = {Journal of Biological Inorganic Chemistry}, volume = {2}, number = {5}, year = {1997}, note = {

Times Cited: 14 Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 15

}, pages = {611-622}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s007750050176}, url = {://WOS:A1997YF71200007}, author = {Archer, M. and Banci, L. and Dikaya, E. and Romao, M. J.} } @article {7280, title = {The crystal structures of two spermadhesins reveal the CUB domain fold}, journal = {Nature Structural Biology}, volume = {4}, number = {10}, year = {1997}, note = {

Times Cited: 110 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; 114

}, pages = {783-788}, type = {Journal Article}, abstract = {n/a}, issn = {1072-8368}, doi = {10.1038/nsb1097-783}, url = {://WOS:A1997YA20300007}, author = {Romero, A. and Romao, M. J. and Varela, P. F. and Kolln, I. and Dias, J. M. and Carvalho, A. L. and Sanz, L. and TopferPetersen, E. and Calvete, J. J.} } @article {7209, title = {Crystallization and preliminary x-ray diffraction studies of aSFP, a bovine seminal plasma protein with a single CUB domain architecture}, journal = {Protein Science}, volume = {6}, number = {3}, year = {1997}, note = {

Times Cited: 7 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; 7

}, pages = {725-727}, type = {Journal Article}, abstract = {n/a}, issn = {0961-8368}, url = {://WOS:A1997WM74000023}, author = {Dias, J. M. and Carvalho, A. L. and Kolln, I. and Calvete, J. J. and TopferPetersen, E. and Varela, P. F. and Romero, A. and Urbanke, C. and Romao, M. J.} } @article {7275, title = {The molybdenum site in the xanthine oxidase-related aldehyde oxidoreductase from Desulfovibrio gigas and a catalytic mechanism for this class of enzymes}, journal = {Journal of Biological Inorganic Chemistry}, volume = {2}, number = {6}, year = {1997}, note = {

Times Cited: 23 Roesch, Notker/C-1182-2010; Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 23

}, pages = {782-785}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s007750050195}, url = {://WOS:000071271700013}, author = {Romao, M. J. and Rosch, N. and Huber, R.} } @article {7303, title = {Prediction of alternative structures of the molybdenum site in the xanthine oxidase-related aldehyde oxide reductase}, journal = {Journal of the American Chemical Society}, volume = {119}, number = {13}, year = {1997}, note = {

Times Cited: 33 Gemming, Sibylle/C-6898-2009; Roesch, Notker/C-1182-2010; Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013; Neyman, Konstantin/F-4055-2011 Romao, Maria/0000-0002-3004-0543; Neyman, Konstantin/0000-0002-5242-5567 33

}, pages = {3159-3160}, type = {Journal Article}, abstract = {n/a}, issn = {0002-7863}, doi = {10.1021/ja963901+}, url = {://WOS:A1997WR15000024}, author = {Voityuk, A. A. and Albert, K. and Kostlmeier, S. and Nasluzov, V. A. and Neyman, K. M. and Hof, P. and Huber, R. and Romao, M. J. and Rosch, N.} } @article {7273, title = {Structure and function of molybdopterin containing enzymes}, journal = {Progress in Biophysics \& Molecular Biology}, volume = {68}, number = {2-3}, year = {1997}, note = {

Times Cited: 68 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 68

}, pages = {121-144}, type = {Journal Article}, abstract = {n/a}, issn = {0079-6107}, url = {://WOS:000074129700001}, author = {Romao, M. J. and Knablein, J. and Huber, R. and Moura, J. J. G.} } @article {7281, title = {The three-dimensional structure of mammalian spermadhesins determined by x-ray crystallography}, journal = {European Journal of Cell Biology}, volume = {74}, year = {1997}, note = {

Times Cited: 0 Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 46 0

}, pages = {13-13}, type = {Journal Article}, abstract = {n/a}, issn = {0171-9335}, url = {://WOS:A1997YA74100013}, author = {Romero, A. and Varela, P. F. and Romao, M. J. and Sanz, L. and TopferPetersen, E. and Calvete, J. J.} } @article {7247, title = {Analysis, design and engineering of simple iron-sulfur proteins: Tales from rubredoxin and desulforedoxin}, journal = {Comments on Inorganic Chemistry}, volume = {19}, number = {1}, year = {1996}, note = {

Times Cited: 10 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; A 10

}, pages = {47-+}, type = {Journal Article}, abstract = {n/a}, issn = {0260-3594}, doi = {10.1080/02603599608032727}, url = {://WOS:A1996WE58400003}, author = {Moura, J. J. G. and Goodfellow, B. J. and Romao, M. J. and Rusnak, F. and Moura, I.} } @article {7279, title = {Crystal structure of flavodoxin from Desulfovibrio desulfuricans ATCC 27774 in two oxidation states}, journal = {European Journal of Biochemistry}, volume = {239}, number = {1}, year = {1996}, note = {

Times Cited: 31 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 32

}, pages = {190-196}, type = {Journal Article}, abstract = {n/a}, issn = {0014-2956}, doi = {10.1111/j.1432-1033.1996.0190u.x}, url = {://WOS:A1996UV76300027}, author = {Romero, A. and Caldeira, J. and Legall, J. and Moura, I. and Moura, J. J. G. and Romao, M. J.} } @article {7307, title = {Crystallographic and fluorescence studies of ligand binding to N-carbamoylsarcosine amidohydrolase from Arthrobacter sp}, journal = {Journal of Molecular Biology}, volume = {263}, number = {2}, year = {1996}, note = {

Times Cited: 18 Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 20

}, pages = {269-283}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1006/jmbi.1996.0574}, url = {://WOS:A1996VP73900015}, author = {Zajc, A. and Romao, M. J. and Turk, B. and Huber, R.} } @article {7233, title = {Protein-crystal density by volume measurement and amino-acid analysis}, journal = {Journal of Applied Crystallography}, volume = {29}, year = {1996}, note = {

Times Cited: 12 Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 4 12

}, pages = {311-317}, type = {Journal Article}, abstract = {n/a}, issn = {0021-8898}, doi = {10.1107/s0021889895014932}, url = {://WOS:A1996VF65000002}, author = {Kiefersauer, R. and Stetefeld, J. and GomisRuth, F. X. and Romao, M. J. and Lottspeich, F. and Huber, R.} } @article {7227, title = {The solution structure of desulforedoxin, a simple iron-sulfur protein - An NMR study of the zinc derivative}, journal = {Journal of Biological Inorganic Chemistry}, volume = {1}, number = {4}, year = {1996}, note = {

Times Cited: 13 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; Pedro, Tavares/B-3654-2008; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Pedro, Tavares/0000-0002-7398-2661; Moura, Isabel/0000-0003-0971-4977; 14

}, pages = {341-354}, type = {Journal Article}, abstract = {n/a}, issn = {0949-8257}, doi = {10.1007/s007750050062}, url = {://WOS:A1996VV70600010}, author = {Goodfellow, B. J. and Tavares, P and Romao, M. J. and Czaja, C. and Rusnak, F. and Legall, J. and Moura, I. and Moura, J. J. G.} } @article {7232, title = {A structure-based catalytic mechanism for the xanthine oxidase family of molybdenum enzymes}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {93}, number = {17}, year = {1996}, note = {

Times Cited: 194 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; Szaleniec, Maciej/A-1198-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 200

}, pages = {8846-8851}, type = {Journal Article}, abstract = {n/a}, issn = {0027-8424}, doi = {10.1073/pnas.93.17.8846}, url = {://WOS:A1996VD43400012}, author = {Huber, R. and Hof, P. and Duarte, R. O. and Moura, J. J. G. and Moura, I. and Liu, M. Y. and Legall, J. and Hille, R. and Archer, M. and Romao, M. J.} } @article {7176, title = {CRYSTAL-STRUCTURE OF DESULFOREDOXIN FROM DESULFOVIBRIO-GIGAS DETERMINED AT 1.8 ANGSTROM RESOLUTION - A NOVEL NONHEME IRON PROTEIN-STRUCTURE}, journal = {Journal of Molecular Biology}, volume = {251}, number = {5}, year = {1995}, note = {

Times Cited: 70 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; Pedro, Tavares/B-3654-2008; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Pedro, Tavares/0000-0002-7398-2661; Moura, Isabel/0000-0003-0971-4977; 71

}, pages = {690-702}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1006/jmbi.1995.0465}, url = {://WOS:A1995RR83900008}, author = {Archer, M. and Huber, R. and Tavares, P and Moura, I. and Moura, J. J. G. and Carrondo, M. A. and Sieker, L. C. and Legall, J. and Romao, M. J.} } @article {7267, title = {CRYSTAL-STRUCTURE OF THE XANTHINE OXIDASE-RELATED ALDEHYDE OXIDOREDUCTASE FROM D-GIGAS}, journal = {Science}, volume = {270}, number = {5239}, year = {1995}, note = {

Times Cited: 371 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 374

}, pages = {1170-1176}, type = {Journal Article}, abstract = {n/a}, issn = {0036-8075}, url = {://WOS:A1995TE90500047}, author = {Romao, M. J. and Archer, M. and Moura, I. and Moura, J. J. G. and Legall, J. and Engh, R. and Schneider, M. and Hof, P. and Huber, R.} } @article {7217, title = {CYTOCHROME-C(6) FROM THE GREEN-ALGA MONORAPHIDIUM-BRAUNII - CRYSTALLIZATION AND PRELIMINARY DIFFRACTION STUDIES}, journal = {Acta Crystallographica Section D-Biological Crystallography}, volume = {51}, year = {1995}, note = {

Times Cited: 6 Matias, Pedro/B-7180-2008; Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Matias, Pedro/0000-0001-6170-451X; Romao, Maria/0000-0002-3004-0543; 2 6

}, pages = {232-234}, type = {Journal Article}, abstract = {n/a}, issn = {0907-4449}, doi = {10.1107/s0907444994005184}, url = {://WOS:A1995QT47100013}, author = {Frazao, C. and Dias, J. M. and Matias, P. M. and Romao, M. J. and Carrondo, M. A. and Hervas, M. and Navarro, J. A. and Delarosa, M. and Sheldrick, G. M.} } @article {7295, title = {MOLECULAR-CLONING AND SEQUENCE-ANALYSIS OF THE GENE OF THE MOLYBDENUM-CONTAINING ALDEHYDE OXIDOREDUCTASE OF DESULFOVIBRIO-GIGAS - THE DEDUCED AMINO-ACID-SEQUENCE SHOWS SIMILARITY TO XANTHINE DEHYDROGENASE}, journal = {European Journal of Biochemistry}, volume = {220}, number = {3}, year = {1994}, note = {

Times Cited: 45 Devreese, bart/B-2011-2009; Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 46

}, pages = {901-910}, type = {Journal Article}, abstract = {n/a}, issn = {0014-2956}, doi = {10.1111/j.1432-1033.1994.tb18693.x}, url = {://WOS:A1994NB53000028}, author = {Thoenes, U. and Flores, O. L. and Neves, A. and Devreese, B. and Van Beeumen, J. J. and Huber, R. and Romao, M. J. and Legall, J. and Moura, J. J. G. and Rodriguespousada, C.} } @article {7268, title = {SUBUNIT COMPOSITION, CRYSTALLIZATION AND PRELIMINARY CRYSTALLOGRAPHIC STUDIES OF THE DESULFOVIBRIO-GIGAS ALDEHYDE OXIDOREDUCTASE CONTAINING MOLYBDENUM AND 2FE-2S CENTERS}, journal = {European Journal of Biochemistry}, volume = {215}, number = {3}, year = {1993}, note = {

Times Cited: 24 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 24

}, pages = {729-732}, type = {Journal Article}, abstract = {n/a}, issn = {0014-2956}, doi = {10.1111/j.1432-1033.1993.tb18085.x}, url = {://WOS:A1993LR70200025}, author = {Romao, M. J. and Barata, B. A. S. and Archer, M. and Lobeck, K. and Moura, I. and Carrondo, M. A. and Legall, J. and Lottspeich, F. and Huber, R. and Moura, J. J. G.} } @article {7276, title = {CRYSTAL-STRUCTURE ANALYSIS, REFINEMENT AND ENZYMATIC-REACTION MECHANISM OF N-CARBAMOYLSARCOSINE AMIDOHYDROLASE FROM ARTHROBACTER SP AT 2.0-ANGSTROM RESOLUTION}, journal = {Journal of Molecular Biology}, volume = {226}, number = {4}, year = {1992}, note = {

Times Cited: 32 Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 34

}, pages = {1111-1130}, type = {Journal Article}, abstract = {n/a}, issn = {0022-2836}, doi = {10.1016/0022-2836(92)91056-u}, url = {://WOS:A1992JK69700018}, author = {Romao, M. J. and Turk, D. and GomisRuth, F. X. and Huber, R. and Schumacher, G. and Mollering, H. and Russmann, L.} } @article {7236, title = {ESCHERICHIA-COLI DIHYDRODIPICOLINATE SYNTHASE - IDENTIFICATION OF THE ACTIVE-SITE AND CRYSTALLIZATION}, journal = {Biochemical Journal}, volume = {288}, year = {1992}, note = {

Times Cited: 64 Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 2 67

}, pages = {691-695}, type = {Journal Article}, abstract = {n/a}, issn = {0264-6021}, url = {://WOS:A1992KB76600052}, author = {Laber, B. and GomisRuth, F. X. and Romao, M. J. and Huber, R.} }