Publications

Export 207 results:
Sort by: [ Author  (Asc)] Title Type Year
A B C D E F G H I J K L M N O P Q R S T U V W X Y Z 
R
Romao, MJ.  2006.  The role of molybdenum in biology. Metal Ions in Biology and Medicine, Vol 9. 9(Alpoim, M.C., Morais, P.V., Eds.).:507-510. Abstract
n/a
Romao, MJ, Kolln I, Dias JM, Carvalho AL, Romero A, Varela PF, Sanz L, Topfer-Petersen E, Calvete JJ.  1997.  Crystal structure of acidic seminal fluid protein (aSFP) at 1.9 angstrom resolution: a bovine polypeptide of the spermadhesin family. Journal of Molecular Biology. 274:650-660., Number 4 AbstractWebsite
n/a
Romao, MJ, Huber R.  1998.  Structure and function of the xanthine-oxidase family of molybdenum enzymes. Metal Sites in Proteins and Models. 90:69-95. AbstractWebsite
n/a
Romao, MJ, Barata BAS, Archer M, Lobeck K, Moura I, Carrondo MA, Legall J, Lottspeich F, Huber R, Moura JJG.  1993.  SUBUNIT COMPOSITION, CRYSTALLIZATION AND PRELIMINARY CRYSTALLOGRAPHIC STUDIES OF THE DESULFOVIBRIO-GIGAS ALDEHYDE OXIDOREDUCTASE CONTAINING MOLYBDENUM AND 2FE-2S CENTERS. European Journal of Biochemistry. 215:729-732., Number 3 AbstractWebsite
n/a
Romão, MJ, Coelho C, Santos-Silva T, Foti A, Terao M, Garattini E, Leimkühler S.  2017.  Structural basis for the role of mammalian aldehyde oxidases in the metabolism of drugs and xenobiotics. Current Opinion in Chemical Biology. 37:39-47. AbstractWebsite

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.

Romao, MJ.  2009.  Molybdenum and tungsten enzymes: a crystallographic and mechanistic overview. Dalton Transactions. :4053-4068., Number 21 AbstractWebsite
n/a
Romao, MJ, Hubert R.  1997.  Crystal structure and mechanism of action of the xanthine oxidase-related aldehyde oxidoreductase from Desulfovibrio gigas. Biochemical Society Transactions. 25:755-757., Number 3 AbstractWebsite
n/a
Romao, MJ, Carvalho AL, Dias JM, Teixeira S, Bourenkov G, Bartunik H, Huber R, Maia L, Mira L.  1999.  Preliminary crystallographic studies of xanthine oxidase purified from rat liver. Journal of Inorganic Biochemistry. 74:281-281., Number 1-4 AbstractWebsite
n/a
Romao, MJ.  2013.  Unraveling new functions and modes of action of molybdenum-dependent enzymes. European Biophysics Journal with Biophysics Letters. 42:S35-S35. AbstractWebsite
n/a
Romao, MJ, Turk D, GomisRuth FX, Huber R, Schumacher G, Mollering H, Russmann L.  1992.  CRYSTAL-STRUCTURE ANALYSIS, REFINEMENT AND ENZYMATIC-REACTION MECHANISM OF N-CARBAMOYLSARCOSINE AMIDOHYDROLASE FROM ARTHROBACTER SP AT 2.0-ANGSTROM RESOLUTION. Journal of Molecular Biology. 226:1111-1130., Number 4 AbstractWebsite
n/a
Romao, MJ, Knablein J, Huber R, Moura JJG.  1997.  Structure and function of molybdopterin containing enzymes. Progress in Biophysics & Molecular Biology. 68:121-144., Number 2-3 AbstractWebsite
n/a
Romao, MJ, Cunha CA, Brondino CD, Moura JJG.  2002.  Molybdenum enzymes in reactions involving aldehydes and acids. Molybdenum and Tungsten: Their Roles in Biological Processes. 39:539-570. AbstractWebsite
n/a
Romao, MJ, Archer M, Moura I, Moura JJG, Legall J, Engh R, Schneider M, Hof P, Huber R.  1995.  CRYSTAL-STRUCTURE OF THE XANTHINE OXIDASE-RELATED ALDEHYDE OXIDOREDUCTASE FROM D-GIGAS. Science. 270:1170-1176., Number 5239 AbstractWebsite
n/a
Romero, A, Varela PF, Romao MJ, Sanz L, TopferPetersen E, Calvete JJ.  1997.  The three-dimensional structure of mammalian spermadhesins determined by x-ray crystallography. European Journal of Cell Biology. 74:13-13. AbstractWebsite
n/a
Romero, A, Caldeira J, Legall J, Moura I, Moura JJG, Romao MJ.  1996.  Crystal structure of flavodoxin from Desulfovibrio desulfuricans ATCC 27774 in two oxidation states. European Journal of Biochemistry. 239:190-196., Number 1 AbstractWebsite
n/a
Romero, A, Romao MJ, Varela PF, Kolln I, Dias JM, Carvalho AL, Sanz L, TopferPetersen E, Calvete JJ.  1997.  The crystal structures of two spermadhesins reveal the CUB domain fold. Nature Structural Biology. 4:783-788., Number 10 AbstractWebsite
n/a
S
Sadeghi, SJ, Valetti F, Cunha CA, Romao MJ, Soares CM, Gilardi G.  2000.  Ionic strength dependence of the non-physiological electron transfer between flavodoxin and cytochrome c(553) from D-vulgaris. Journal of Biological Inorganic Chemistry. 5:730-737., Number 6 AbstractWebsite
n/a
Santarsia, S, Grosso AS, Trovão F, Jiménez-Barbero J, Carvalho AL, Nativi C, Marcelo F.  2018.  Molecular recognition of a Thomsen-Friedenreich antigen mimetic targeting human galectin-3, 2018. ChemMedChem. Aug 9. doi: 10.1002/cmdc.201800525. [Epub ahead of print](ja): Wiley-Blackwell AbstractWebsite

Overexpression of the Thomsen-Friedenreich (TF) antigen in cell membrane proteins occurs in 90% of adenocarcinomas. Additionally, the binding of the TF-antigen to human galectin-3 (Gal-3), also frequently overexpressed in malignancy, promotes cancer progression and metastasis. In this context, structures that interfere with this specific interaction display the potential to prevent cancer metastasis. Herein, a multidisciplinary approach, combining the optimized synthesis of a TF-antigen mimetic with NMR, X-ray crystallography methods and isothermal titration calorimetry assays has been employed to unravel the molecular structural details that govern the Gal-3/TF-mimetic interaction. The TF-mimetic presents a binding affinity for Gal-3 similar to the TF-natural antigen and retains the binding epitope and the bioactive conformation observed for the native antigen. Furthermore, from a thermodynamic perspective a decrease in the enthalpic contribution was observed for the Gal-3/TF-mimetic complex, however this behaviour is compensated by a favourable entropy gain. From a structural perspective, these results establish our TF-mimetic as a scaffold to design multivalent solutions to potentially interfere with Gal-3 aberrant interactions and likely be used to hamper Gal-3-mediated cancer cells adhesion and metastasis.

Santos, MFA, Sciortino G, Correia I, Fernandes ACP, Santos-Silva T, Pisanu F, Garribba E, Pessoa JC.  2022.  Binding of VIVO2+, VIVOL, VIVOL2 and VVO2L Moieties to Proteins: X-ray/Theoretical Characterization and Biological Implications, 2022. Chemistry – A European JournalChemistry – A European Journal. 28(40):e202200105.: John Wiley & Sons, Ltd AbstractWebsite

Abstract Vanadium compounds have frequently been proposed as therapeutics, but their application has been hampered by the lack of information on the different V-containing species that may form and how these interact with blood and cell proteins, and with enzymes. Herein, we report several resolved crystal structures of lysozyme with bound VIVO2+ and VIVOL2+, where L=2,2?-bipyridine or 1,10-phenanthroline (phen), and of trypsin with VIVO(picolinato)2 and VVO2(phen)+ moieties. Computational studies complete the refinement and shed light on the relevant role of hydrophobic interactions, hydrogen bonds, and microsolvation in stabilizating the structure. Noteworthy is that the trypsin?VVO2(phen) and trypsin?VIVO(OH)(phen) adducts correspond to similar energies, thus suggesting a possible interconversion under physiological/biological conditions. The obtained data support the relevance of hydrolysis of VIV and VV complexes in the several types of binding established with proteins and the formation of different adducts that might contribute to their pharmacological action, and significantly widen our knowledge of vanadium?protein interactions.

dos Santos, MMC, Sousa PMP, Goncalves MLS, Romao MJ, Moura I, Moura JJG.  2004.  Direct electrochemistry of the Desulfovibrio gigas aldehyde oxidoreductase. European Journal of Biochemistry. 271:1329-1338., Number 7 AbstractWebsite
n/a
dos Santos, R, Romão MJ, Roque ACA, Carvalho AL.  2021.  Magnetic particles used in a new approach for designed protein crystallization. CrystEngComm. 23:1083-1090.: The Royal Society of Chemistry AbstractWebsite

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.

dos Santos, R, Iria I, Manuel AM, Leandro AP, Madeira CAC, Goncalves J, Carvalho AL, Roque AC.  2020.  Magnetic Precipitation: A New Platform for Protein Purification, 2020. Biotechnology JournalBiotechnology Journal. n/a(n/a):2000151.: John Wiley & Sons, Ltd AbstractWebsite

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°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

Santos, MFA, Seixas JD, Coelho AC, Mukhopadhyay A, Reis PM, Romao MJ, Romao CC, Santos-Silva T.  2012.  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 of Inorganic Biochemistry. 117:285-291. AbstractWebsite
n/a
Santos, MFA, Correia I, Oliveira AR, Garribba E, Pessoa JC, Santos-Silva T.  2014.  Vanadium Complexes as Prospective Therapeutics: Structural Characterization of a VIV Lysozyme Adduct. European Journal of Inorganic Chemistry. :n/a–n/a.: WILEY-VCH Verlag AbstractWebsite

The biological activity of vanadium complexes, namely, as insulin enhancers, is well known. We report a combined X-ray crystallography, electron paramagnetic resonance, and density functional theory study of the interaction of vanadium picolinate complexes with hen egg white lysozyme (HEWL). We show that the VIVO(pic)2 complex covalently binds to the COO– group of the side chain of Asp52 of HEWL. The long VIV=O bond obtained in the X-ray study is explained to be due to reduction of VIV to VIII during exposure of the crystals to the intense X-ray beam.