@inbook {9911, title = {Nitrite biosensing using cytochrome c nitrite reductase: Towards a disposable strip electrode}, booktitle = {Biomedical Engineering Systems and Technologies, Communications in Computer and Information Science}, year = {2013}, pages = {41-51}, publisher = {Springer, Heidelberg}, organization = {Springer, Heidelberg}, address = {DOI:10.1007/978-3-642-38256-7_3}, author = {Correia C. and Rodrigues M. and Silveira C.M. and Moura J.J.G. and Ochoteco E. and Jubete E. and Almeida M.G.} } @inbook {16121, title = {Carbon dioxide utilisation - bioelectrochemical approaches}, booktitle = {Enzymes for Solving Humankind{\textquoteright}s Problems, Moura J.J.G., Moura I., Maia L.B. (eds)}, year = {2021}, pages = {83-108}, author = {Cordas C.M. and Moura J.J.G. and Escapa A. and Mateos R.} } @article {2538, title = {Primary sequence, oxidation-reduction potentials and tertiary-structure prediction of Desulfovibrio desulfuricans ATCC 27774 flavodoxin}, journal = {Eur J Biochem}, volume = {220}, number = {3}, year = {1994}, note = {

0014-2956 (Print)0014-2956 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {Mar 15}, pages = {987-95}, abstract = {

Flavodoxin was isolated and purified from Desulfovibrio desulfuricans ATCC 27774, a sulfate-reducing organism that can also utilize nitrate as an alternative electron acceptor. Mid-point oxidation-reduction potentials of this flavodoxin were determined by ultraviolet/visible and EPR methods coupled to potentiometric measurements and their pH dependence studied in detail. The redox potential E2, for the couple oxidized/semiquinone forms at pH 6.7 and 25 degrees C is -40 mV, while the value for the semiquinone/hydroquinone forms (E1), at the same pH, -387 mV. E2 varies linearly with pH, while E1 is independent of pH at high values. However, at low pH (\< 7.0), this value is less negative, compatible with a redox-linked protonation of the flavodoxin hydroquinone. A comparative study is presented for Desulfovibrio salexigens NCIB 8403 flavodoxin [Moura, I., Moura, J.J.G., Bruschi, M. \& LeGall, J. (1980) Biochim. Biophys. Acta 591, 1-8]. The complete primary amino acid sequence was obtained by automated Edman degradation from peptides obtained by chemical and enzymic procedures. The amino acid sequence was confirmed by FAB/MS. Using the previously determined tridimensional structure of Desulfovibrio vulgaris flavodoxin as a model [similarity, 48.6\%; Watenpaugh, K.D., Sieker, L.C., Jensen, L.H., LeGall, J. \& Dubourdieu M. (1972) Proc. Natl Acad. Sci. USA 69, 3185-3188], the tridimensional structure of D. desulfuricans ATCC 27774 flavodoxin was predicted using AMBER force-field calculations.

}, keywords = {Amino Acid Sequence, Base Sequence, Desulfovibrio/*chemistry, Electron Spin Resonance Spectroscopy, Flavodoxin/*chemistry, Hydrogen-Ion Concentration, Molecular Sequence Data, Oxidation-Reduction, Protein Structure, Tertiary, Sequence Homology, Amino Acid, Spectrophotometry, Ultraviolet, Spectrum Analysis}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=8143752 }, author = {Caldeira, J. and Palma, P. N. and Regalla, M. and Lampreia, J. and Calvete, J. and Schafer, W. and Legall, J. and Moura, I. and Moura, J. J.} } @article {2443, title = {Analysis of the electron paramagnetic resonance properties of the [2Fe-2S]1+ centers in molybdenum enzymes of the xanthine oxidase family: assignment of signals I and II}, journal = {Biochemistry}, volume = {39}, number = {10}, year = {2000}, note = {

0006-2960 (Print)0006-2960 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {Mar 14}, pages = {2700-7}, abstract = {

Molybdoenzymes of the xanthine oxidase family contain two [2Fe-2S](1+,2+) clusters that are bound to the protein by very different cysteine motifs. In the X-ray crystal structure of Desulfovibrio gigas aldehyde oxidoreductase, the cluster ligated by a ferredoxin-type motif is close to the protein surface, whereas that ligated by an unusual cysteine motif is in contact with the molybdopterin [Romao, M. J., Archer, M., Moura, I., Moura, J. J. G., LeGall, J., Engh, R., Schneider, M., Hof, P., and Huber, R. (1995) Science 270, 1170-1176]. These two clusters display distinct electron paramagnetic resonance (EPR) signals: the less anisotropic one, called signal I, is generally similar to the g(av) approximately 1.96-type signals given by ferredoxins, whereas signal II often exhibits anomalous properties such as very large g values, broad lines, and very fast relaxation properties. A detailed comparison of the temperature dependence of the spin-lattice relaxation time and of the intensity of these signals in D. gigas aldehyde oxidoreductase and in milk xanthine oxidase strongly suggests that the peculiar EPR properties of signal II arise from the presence of low-lying excited levels reflecting significant double exchange interactions. The issue raised by the assignment of signals I and II to the two [2Fe-2S](1+) clusters was solved by using the EPR signal of the Mo(V) center as a probe. The temperature dependence of this signal could be quantitatively reproduced by assuming that the Mo(V) center is coupled to the cluster giving signal I in xanthine oxidase as well as in D. gigas aldehyde oxidoreductase. This demonstrates unambiguously that, in both enzymes, signal I arises from the center which is closest to the molybdenum cofactor.

}, keywords = {Aldehyde Oxidoreductases/chemistry, Animals, Cattle, Desulfovibrio/enzymology, Electron Spin Resonance Spectroscopy, Iron-Sulfur Proteins/*chemistry, Liver/enzymology, Metalloproteins/chemistry, Milk/enzymology, Molybdenum/*chemistry, Rabbits, Spin Labels, Temperature, Turkeys, Xanthine Oxidase/*chemistry}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=10704221 }, author = {Caldeira, J. and Belle, V. and Asso, M. and Guigliarelli, B. and Moura, I. and Moura, J. J. and Bertrand, P.} } @article {2498, title = {EPR and Mossbauer spectroscopic studies on enoate reductase}, journal = {J Biol Chem}, volume = {271}, number = {31}, year = {1996}, note = {

0021-9258 (Print)0021-9258 (Linking)Journal Article

}, month = {Aug 2}, pages = {18743-8}, abstract = {

Enoate reductase (EC 1.3.1.31) is a protein isolated from Clostridium tyrobutyricum that contains iron, labile sulfide, FAD, and FMN. The enzyme reduces the alpha,beta carbon-carbon double bond of nonactivated 2-enoates and in a reversible way that of 2-enals at the expense of NADH or reduced methyl viologen. UV-visible and EPR potentiometric titrations detect a semiquinone species in redox intermediate states characterized by an isotropic EPR signal at g = 2.0 without contribution at 580 nm. EPR redox titration shows two widely spread mid-point redox potentials (-190 and -350 mV at pH 7. 0), and a nearly stoichiometric amount of this species is detected. The data suggest the semiquinone radical has an anionic nature. In the reduced form, the [Fe-S] moiety is characterized by a single rhombic EPR spectrum, observed in a wide range of temperatures (4. 2-60 K) with g values at 2.013, 1.943, and 1.860 (-180 mV at pH 7.0). The gmax value is low when compared with what has been reported for other iron-sulfur clusters. Mossbauer studies reveal the presence of a [4Fe-4S]+2/+1 center. One of the subcomponents of the spectrum shows an unusually large value of quadrupole splitting (ferrous character) in both the oxidized and reduced states. Substrate binding to the reduced enzyme induces subtle changes in the spectroscopic Mossbauer parameters. The Mossbauer data together with known kinetic information suggest the involvement of this iron-sulfur center in the enzyme mechanism.

}, keywords = {*Oxidoreductases Acting on CH-CH Group Donors, Binding Sites, Catalysis, Clostridium/enzymology, Electron Spin Resonance Spectroscopy, Iron-Sulfur Proteins/chemistry/metabolism, Iron/chemistry, Kinetics, Molecular Structure, Oxidation-Reduction, Oxidoreductases/*chemistry/metabolism, Spectrophotometry, Spectroscopy, Mossbauer, Substrate Specificity, Sulfur/chemistry, Temperature}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=8702530 }, author = {Caldeira, J. and Feicht, R. and White, H. and Teixeira, M. and Moura, J. J. and Simon, H. and Moura, I.} } @article {2668, title = {Spectroscopic studies of the oxidation-reduction properties of three forms of ferredoxin from Desulphovibrio gigas}, journal = {Biochim Biophys Acta}, volume = {490}, number = {2}, year = {1977}, note = {

0006-3002 (Print)0006-3002 (Linking)Journal Article

}, month = {Feb 22}, pages = {311-21}, abstract = {

Electron paramagnetic resonance spectra were recorded of three forms of Desulphovibrio gigas ferredoxin, FdI, FdI{\textquoteright} and FdII. The g = 1.94 signal seen in dithionite-reduced samples is strong in FdI, weaker in FdI{\textquoteright} and very small in FdII. The g = 2.02 signal in the oxidized proteins is weak in FdI and strongest in FdII. It is concluded that most of the 4Fe-4S centres in FdI change between states C- and C2-; FdI{\textquoteright} contain both types of centre. There is no evidence that any particular centre can change reversibly between all three oxidation states. Circular dichroism spectra show differences between FdI and FdII even in the diamagnetic C2- state. The redox potentials of the iron-sulphur centres of the three oligomers (forms) are different. After formation of the apo-protein of FdII and reconstitution with iron and sulphide, the protein behaves more like FdI, showing a strong g = 1.94 signal in the reduced states.

}, keywords = {*Ferredoxins/metabolism, Binding Sites, Circular Dichroism, Desulfovibrio/*metabolism, Dithionite, Electron Spin Resonance Spectroscopy, Oxidation-Reduction, Protein Binding, Protein Conformation}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=189829 }, author = {Cammack, R. and Rao, K. K. and Hall, D. O. and Moura, J. J. and Xavier, A. V. and Bruschi, M. and Legall, J. and Deville, A. and Gayda, J. P.} } @article {2623, title = {ESR studies of cytochrome c3 from Desulfovibrio desulfuricans strain Norway 4: Midpoint potentials of the four haems, and interactions with ferredoxin and colloidal sulphur}, journal = {Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology}, volume = {784}, number = {1}, year = {1984}, note = {n/a}, pages = {68-74}, type = {doi: 10.1016/0167-4838(84)90174-2}, abstract = {n/a}, keywords = {(D. desulfuricans), Cytochrome c3, ESR, Heme redox potential}, isbn = {0167-4838}, url = {http://www.sciencedirect.com/science/article/pii/0167483884901742}, author = {Cammack, R. and Fauque, G. and Moura, J. J. G. and Legall, J.} } @article {10863, title = {Orange protein from Desulfovibrio alaskensis G20: insights into the Mo-Cu cluster protein-assisted synthesis}, journal = {J Biol Inorg Chem}, volume = {21}, year = {2016}, pages = {53-62}, author = {Carepo, M. S. and Carreira, C. and Grazina, R. and M.E. Zakrzewska and Dolla, A. and Aubert, C. and Pauleta, S. R. and Moura, J. J. G. and Moura, I.} } @article {2414, title = {17O ENDOR detection of a solvent-derived Ni-(OH(x))-Fe bridge that is lost upon activation of the hydrogenase from Desulfovibrio gigas}, journal = {J Am Chem Soc}, volume = {124}, number = {2}, year = {2002}, note = {

0002-7863 (Print)0002-7863 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}tResearch Support, U.S. Gov{\textquoteright}t, Non-P.H.S.

}, month = {Jan 16}, pages = {281-6}, abstract = {

Crystallographic studies of the hydrogenases (Hases) from Desulfovibrio gigas (Dg) and Desulfovibrio vulgaris Miyazaki (DvM) have revealed heterodinuclear nickel-iron active centers in both enzymes. The structures, which represent the as-isolated (unready) Ni-A (S = (1)/(2)) enzyme state, disclose a nonprotein ligand (labeled as X) bridging the two metals. The bridging atom was suggested to be an oxygenic (O(2)(-) or OH(-)) species in Dg Hase and an inorganic sulfide in DvM Hase. To determine the nature and chemical characteristics of the Ni-X-Fe bridging ligand in Dg Hase, we have performed 35 GHz CW (17)O ENDOR measurements on the Ni-A form of the enzyme, exchanged into H(2)(17)O, on the active Ni-C (S = (1)/(2)) form prepared by H(2)-reduction of Ni-A in H(2)(17)O, and also on Ni-A formed by reoxidation of Ni-C in H(2)(17)O. In the native state of the protein (Ni-A), the bridging ligand does not exchange with the H(2)(17)O solvent. However, after a reduction/reoxidation cycle (Ni-A --\> Ni-C --\> Ni-A), an (17)O label is introduced at the active site, as seen by ENDOR. Detailed analysis of a 2-D field-frequency plot of ENDOR spectra taken across the EPR envelope of Ni-A((17)O) shows that the incorporated (17)O has a roughly axial hyperfine tensor, A((17)O) approximately [5, 7, 20] MHz, discloses its orientation relative to the g tensor, and also yields an estimate of the quadrupole tensor. The substantial isotropic component (a(iso)((17)O) approximately 11 MHz) of the hyperfine interaction indicates that a solvent-derived (17)O is indeed a ligand to Ni and thus that the bridging ligand X in the Ni-A state of Dg Hase is indeed an oxygenic (O(2)(-) or OH(-)) species; comparison with earlier EPR results by others indicates that the same holds for Ni-B. The small (57)Fe hyperfine coupling seen previously for Ni-A (A((57)Fe) approximately 0.9 MHz) is now shown to persist in Ni-C, A((57)Fe) approximately 0.8 MHz. However, the (17)O signal is lost upon reductive activation to the Ni-C state; reoxidation to Ni-A leads to the reappearance of the signal. Consideration of the electronic structure of the EPR-active states of the dinuclear center leads us to suggest that the oxygenic bridge in Ni-A(B) is lost in Ni-C and is re-formed from solvent upon reoxidation to Ni-A. This implies that the reductive activation to Ni-C opens Ni/Fe coordination sites which may play a central role in the enzyme{\textquoteright}s activity.

}, keywords = {Binding Sites, Desulfovibrio/*enzymology, Electron Spin Resonance Spectroscopy/methods, Hydrogenase/*chemistry/*metabolism, Iron/chemistry/metabolism, Models, Molecular, Nickel/chemistry/metabolism, Oxygen Isotopes}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=11782180 }, author = {Carepo, M. and Tierney, D. L. and Brondino, C. D. and Yang, T. C. and Pamplona, A. and Telser, J. and Moura, I. and Moura, J. J. and Hoffman, B. M.} } @article {2415, title = {Hydrogen metabolism in Desulfovibrio desulfuricans strain New Jersey (NCIMB 8313)--comparative study with D. vulgaris and D. gigas species}, journal = {Anaerobe}, volume = {8}, number = {6}, year = {2002}, note = {

1075-9964 (Print)1075-9964 (Linking)Journal Article

}, month = {Dec}, pages = {325-32}, abstract = {

This article aims to study hydrogen production/consumption in Desulfovibrio (D.) desulfuricans strain New Jersey, a sulfate reducer isolated from a medium undergoing active biocorrosion and to compare its hydrogen metabolism with two other Desulfovibrio species, D. gigas and D. vulgaris Hildenborough. Hydrogen production was followed during the growth of these three bacterial species under different growth conditions: no limitation of sulfate and lactate, sulfate limitation, lactate limitation, pyruvate/sulfate medium and in the presence of molybdate. Hydrogen production/consumption by D. desulfuricans shows a behavior similar to that of D. gigas but a different one from that of D. vulgaris, which produces higher quantities of hydrogen on lactate/sulfate medium. The three species are able to increase the hydrogen production when the sulfate became limiting. Moreover, in a pyruvate/sulfate medium hydrogen production was lower than on lactate/sulfate medium. Hydrogen production by D. desulfuricans in presence of molybdate is extremely high. Hydrogenases are key enzymes on production/consumption of hydrogen in sulfate reducing organisms. The specific activity, number and cellular localization of hydrogenases vary within the three Desulfovibrio species used in this work, which could explain the differences observed on hydrogen utilization.

}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=16887677 }, author = {Carepo, M. and Baptista, J. F. and Pamplona, A. and Fauque, G. and Moura, J. J. and Reis, M. A.} } @article {6383, title = {Mo-Cu metal cluster formation and binding in an orange protein isolated from Desulfovibrio gigas}, journal = {J Biol Inorg Chem}, volume = {19}, year = {2014}, pages = {605-614}, author = {Carepo, M. S. and Pauleta, S. R. and A.G. Wedd and Moura, J. J. G. and Moura, I.} } @article {14411, title = {Genomic organization, gene expression and activity profile of Marinobacter hydrocarbonoclasticus denitrification enzymes}, journal = {PEERJ}, volume = {6}, year = {2018}, pages = {DOI: 10.7717/peerj.5603}, author = {Carreira, C. and Mestre, O. and Nunes, R.F. and Moura, I. and Pauleta, S. R.} } @article {2303, title = {Can ultrasonic energy efficiently speed (18)O-labeling of proteins?}, journal = {Proteomics}, volume = {9}, number = {21}, year = {2009}, note = {

Times Cited: 6

}, month = {Nov}, pages = {4974-4977}, abstract = {

We report in this work on the robustness of ultrasonic energy as a tool to speed the isotopic labeling of proteins using the (18)O-decoupling procedure. The first part of the decoupling procedure, comprising protein denaturation, reduction, alkylation and digestion, is done in 8 min under the effects of an ultrasonic field whilst the second part, the isotopic labeling, was assayed with and without the use of ultrasonic energy. Our results clearly demonstrate that the (18)O-isotopic labeling in a decoupling procedure cannot be accelerated using an ultrasonic field.

}, isbn = {1615-9853}, url = {http://dx.doi.org/10.1002/pmic.200900197}, author = {Carreira, Ricardo J. and Lodeiro, Carlos and Diniz, Mario S. and Moura, Isabel and Capelo, Jose L.} } @article {15899, title = {The effect of pH on Marinobacter hydrocarbonoclasticus denitrification pathway and nitrous oxide reductase}, journal = {J Biol Inorg Chem}, volume = {25}, year = {2020}, pages = {927}, author = {Carreira, C. and Nunes, R.F. and Mestre, O. and Moura, I. and Pauleta, S. R.} } @article {2341, title = {New findings for in-gel digestion accelerated by high-intensity focused ultrasound for protein identification by matrix-assisted laser desorption ionization time-of-flight mass spectrometry}, journal = {Journal of Chromatography A}, volume = {1153}, number = {1-2}, year = {2007}, note = {

Times Cited: 15

}, month = {Jun 15}, pages = {291-299}, abstract = {

New findings in sample treatment based on high-intensity focused ultrasound (HIFU) for protein digestion after polyacrylamide gel electrophoresis separation are presented. The following variables were studied: (i) sample volume; (ii) sonotrode diameter; (iii) previous protein denaturation; (iv) cooling; (v) enzyme concentration; and (vi) protein concentration. Results showed that positive protein identification could be done after protein separation by gel electrophoresis through peptide mass fingerprint (PMF) in a volume as low as 25 mu L. The time needed was less than 2 min and no cooling was necessary. The importance of the sonotrode diameter was negligible. On the other hand, protein denaturation before sonication was a trade-off for the success of procedure here described. The protein coverage was raised from 5 to 30\%, and the number of peptides matching the proteins was also increased in a percentage ranging 10-100\% when the classical overnight treatment is compared with the proposed HIFU procedure. The minimum amount of protein that can be identified using the HIFU sample treatment by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was 0.06 mu g. The lower concentration of trypsin successfully used to obtain an adequate protein digestion was 3.6 mu g/mL. (c) 2006 Elsevier B.V. All rights reserved.

}, isbn = {0021-9673}, url = {http://dx.doi.org/10.1016/j.chroma.2006.09.078}, author = {Carreira, R. J. and Cordeiro, F. M. and Moro, A. J. and Rivas, M. G. and Rial-Otero, R. and Gaspar, E. M. and Moura, I. and Capelo, J. L.} } @article {2304, title = {Can ultrasonic energy efficiently speed (18)O-labeling of proteins?}, journal = {Proteomics}, volume = {9}, number = {21}, year = {2009}, note = {

Times Cited: 6

}, month = {Nov}, pages = {4974-4977}, abstract = {

We report in this work on the robustness of ultrasonic energy as a tool to speed the isotopic labeling of proteins using the (18)O-decoupling procedure. The first part of the decoupling procedure, comprising protein denaturation, reduction, alkylation and digestion, is done in 8 min under the effects of an ultrasonic field whilst the second part, the isotopic labeling, was assayed with and without the use of ultrasonic energy. Our results clearly demonstrate that the (18)O-isotopic labeling in a decoupling procedure cannot be accelerated using an ultrasonic field.

}, isbn = {1615-9853}, url = {http://dx.doi.org/10.1002/pmic.200900197}, author = {Carreira, Ricardo J. and Lodeiro, Carlos and Diniz, Mario S. and Moura, Isabel and Capelo, Jose L.} } @article {12429, title = {The catalytic cycle of nitrous oxide reductase - The enzyme that catalyzes the last step of denitrification}, journal = {J Inorg Biochem}, volume = {177}, year = {2017}, pages = {423-434}, author = {Carreira, C. and Pauleta, S. R. and Moura, I.} } @article {17740, title = {Electrode Kinetics of Ion Jelly and Ion Sol-Gel Redox Materials on Screen-Printed Electrodes}, journal = {Appl Sci}, volume = {12}, year = {2022}, pages = {2087}, author = {R.N.H. Carvalho and Cordas, C. M. and Fonseca, L. P.} } @article {11666, title = {Sandwich-Type Enzymatic Fuel Cell Based on a New Electro-Conductive Material - Ion Jelly}, journal = {Chemistry Select}, volume = {1}, year = {2016}, pages = {6546{\textendash}6552}, url = {http://dx.doi.org/10.1002/slct.201601640}, author = {Carvalho, R. and Almeida, R and Moura, J. J. G. and Louren{\c c}o, N. and Fonseca, L. and Cordas, C. M.} } @article {14774, title = {Fluorescence anisotropy of fluorescein varies according to pH: lessons for binding studies}, journal = {J Photochem Photobiol A: Chemistry}, volume = {372}, year = {2019}, pages = {59-62}, author = {Castro, N.S.S. and Laia, C. A. T. and Moura, I. and Carepo, M. S.} } @article {14412, title = {Small phospho-donors phosphorylate MorR without inducing protein conformational changes}, journal = {Biophys Chem}, volume = {240}, year = {2018}, pages = {25-33}, author = {Castro, N.S.S. and Laia, C. A. T. and B.K. Maiti and Cerqueira, N. and Moura, I. and Carepo, M. S. P.} } @article {8442, title = {Periplasmic nitrate reductase and formate dehydrogenase: similar molecular architectures with very different enzymatic activities}, journal = {Acc Chem Res}, volume = {48}, year = {2015}, pages = {2875-2884}, author = {Cerqueira, N. and Gonzalez, P. J. and Fernandes, P. A. and Moura, J. J. G. and Ramos, M. J.} } @article {5882, title = {The sulfur-shift: an activation mechanism for periplasmic nitrate reductase and formate dehydrogenase}, journal = {Inorg Chem}, volume = {52}, year = {2013}, pages = {10766-10772}, author = {Cerqueira, N. and Fernandes, P. and Gonz{\'a}lez, P. and Moura, J. J. G. and Ramos, M. J.} } @article {2301, title = {The effect of the sixth sulfur ligand in the catalytic mechanism of periplasmic nitrate reductase}, journal = {J Comput Chem}, volume = {30}, number = {15}, year = {2009}, note = {

1096-987X (Electronic)0192-8651 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {Nov 30}, pages = {2466-84}, abstract = {

The catalytic mechanism of nitrate reduction by periplasmic nitrate reductases has been investigated using theoretical and computational means. We have found that the nitrate molecule binds to the active site with the Mo ion in the +6 oxidation state. Electron transfer to the active site occurs only in the proton-electron transfer stage, where the Mo(V) species plays an important role in catalysis. The presence of the sulfur atom in the molybdenum coordination sphere creates a pseudo-dithiolene ligand that protects it from any direct attack from the solvent. Upon the nitrate binding there is a conformational rearrangement of this ring that allows the direct contact of the nitrate with Mo(VI) ion. This rearrangement is stabilized by the conserved methionines Met141 and Met308. The reduction of nitrate into nitrite occurs in the second step of the mechanism where the two dimethyl-dithiolene ligands have a key role in spreading the excess of negative charge near the Mo atom to make it available for the chemical reaction. The reaction involves the oxidation of the sulfur atoms and not of the molybdenum as previously suggested. The mechanism involves a molybdenum and sulfur-based redox chemistry instead of the currently accepted redox chemistry based only on the Mo ion. The second part of the mechanism involves two protonation steps that are promoted by the presence of Mo(V) species. Mo(VI) intermediates might also be present in this stage depending on the availability of protons and electrons. Once the water molecule is generated only the Mo(VI) species allow water molecule dissociation, and, the concomitant enzymatic turnover.

}, keywords = {Catalysis, Catalytic Domain, Computer Simulation, Ligands, Models, Chemical, Molybdenum/*chemistry, Nitrate Reductase/chemistry/*metabolism, Nitrates/chemistry, Organometallic Compounds/*chemistry, Oxidation-Reduction, Sulfhydryl Compounds/*chemistry}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=19360810 }, author = {Cerqueira, N. M. and Gonzalez, P. J. and Brondino, C. D. and Romao, M. J. and Romao, C. C. and Moura, I. and Moura, J. J.} } @article {2537, title = {Cloning, sequencing and overexpression of the Desulfovibrio gigas ferredoxin gene in E. coli}, journal = {FEBS Lett}, volume = {351}, number = {3}, year = {1994}, note = {

0014-5793 (Print)0014-5793 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}tResearch Support, U.S. Gov{\textquoteright}t, P.H.S.

}, month = {Sep 12}, pages = {401-4}, abstract = {

We have cloned the gene encoding Desulfovibrio gigas ferredoxin using a photodigoxigenin-labelled probe synthesized with the polymerase chain reaction. The DNA sequence of the gene predicts a polypeptide of 58 residues after removal of the initial formyl methionine (polypeptide M(r) = 6,276). The ferredoxin gene was expressed in aerobically grown E. coli behind the lac promoter of pUC18 resulting in a high level of ferredoxin expression which comprises about 10\% of the total cell protein. EPR analysis of recombinant ferredoxin revealed the presence of a [3Fe-4S] cluster which is characteristic of native D. gigas ferredoxin II.

}, keywords = {Amino Acid Sequence, Base Sequence, Cloning, Molecular, Desulfovibrio/*genetics, DNA, Bacterial, Escherichia coli/*genetics, Ferredoxins/*genetics, Molecular Sequence Data, Recombinant Proteins/genetics, Spectrum Analysis}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=8082803 }, author = {Chen, B. and Menon, N. K. and Dervertarnian, L. and Moura, J. J. and Przybyla, A. E.} } @article {2412, title = {Electronic structure description of the mu(4)-sulfide bridged tetranuclear Cu(Z) center in N(2)O reductase}, journal = {J Am Chem Soc}, volume = {124}, number = {5}, year = {2002}, note = {

0002-7863 (Print)0002-7863 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}tResearch Support, U.S. Gov{\textquoteright}t, Non-P.H.S.Research Support, U.S. Gov{\textquoteright}t, P.H.S.

}, month = {Feb 6}, pages = {744-5}, abstract = {

Spectroscopy coupled with density functional calculations has been used to define the spin state, oxidation states, spin distribution, and ground state wave function of the mu4-sulfide bridged tetranuclear CuZ cluster of nitrous oxide reductase. Initial insight into the electronic contribution to N2O reduction is developed, which involves a sigma superexchange pathway through the bridging sulfide.

}, keywords = {Circular Dichroism, Copper/*chemistry, Electron Spin Resonance Spectroscopy, Molecular Structure, Oxidoreductases/*chemistry, Spectrometry, X-Ray Emission/methods, Sulfides/*chemistry}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=11817937 }, author = {Chen, P. and DeBeer George, S. and Cabrito, I. and Antholine, W. E. and Moura, J. J. and Moura, I. and Hedman, B. and Hodgson, K. O. and Solomon, E. I.} } @article {2340, title = {Highly sensitive nitrite biosensor based on the electrical wiring of nitrite reductase by ZnCr-AQS LDH}, journal = {Electrochemistry Communications}, volume = {9}, number = {9}, year = {2007}, note = {

Times Cited: 21

}, month = {Sep}, pages = {2240-2245}, abstract = {

A biosensor for amperometric determination of nitrite was developed using cytochrome c nitrite reductase (ccNiR) from Desulfovibrio desulfuricans immobilized and electrically connected on a glassy carbon electrode by entrapment into redox active [ZnCr-AQS] layered double hydroxide containing anthraquinone-2-sulfonate (AQS). The transduction step corresponded to the electro-enzymatic reduction of nitrite by immobilized AQS molecules at -0.6 V. The biosensor showed a fast response to nitrite (5 s) with a linear range between 0.015 and 2.35 mu M, a sensitivity of 1.8 A M-1 cm(-2) and a detection limit of 4 nM. The apparent Michaelis-Menten constant (K-M(app)) M was 7.5 mu M. (c) 2007 Elsevier B.V. All rights reserved.

}, isbn = {1388-2481}, url = {http://dx.doi.org/10.1016/j.elecom.2007.05.030}, author = {Chen, H. and Mousty, C. and Cosnier, S. and Silveira, C. and Moura, J. J. G. and Almeida, M. G.} } @article {2413, title = {Spectroscopic and electronic structure studies of the mu(4)-sulfide bridged tetranuclear Cu(Z) cluster in N(2)O reductase: molecular insight into the catalytic mechanism}, journal = {J Am Chem Soc}, volume = {124}, number = {35}, year = {2002}, note = {

0002-7863 (Print)0002-7863 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}tResearch Support, U.S. Gov{\textquoteright}t, P.H.S.

}, month = {Sep 4}, pages = {10497-507}, abstract = {

Spectroscopic methods combined with density functional calculations are used to develop a detailed bonding description of the mu(4)-sulfide bridged tetranuclear Cu(Z) cluster in N(2)O reductase. The ground state of Cu(Z) has the 1Cu(II)/3Cu(I) configuration. The single electron hole dominantly resides on one Cu atom (Cu(I)) and partially delocalizes onto a second Cu atom (Cu(II)) via a Cu(I)-S-Cu(II) sigma/sigma superexchange pathway which is manifested by a Cu(II) --\> Cu(I) intervalence transfer transition in absorption. The observed excited-state spectral features of Cu(Z) are dominated by the S --\> Cu(I) charge-transfer transitions and Cu(I) based d-d transitions. The intensity pattern of individual S --\> Cu(I) charge-transfer transitions reflects different bonding interactions of the sulfur valence orbitals with the four Cu{\textquoteright}s in the Cu(Z) cluster, which are consistent with the individual Cu-S force constants obtained from a normal coordinate analysis of the Cu(Z) resonance Raman frequencies and profiles. The Cu(I) d orbital splitting pattern correlates with its distorted T-shaped ligand field geometry and accounts for the observed low g( parallel ) value of Cu(Z) in EPR. The dominantly localized electronic structure description of the Cu(Z) site results from interactions of Cu(II) with the two additional Cu{\textquoteright}s of the cluster (Cu(III)/Cu(IV)), where the Cu-Cu electrostatic interactions lead to hole localization with no metal-metal bonding. The substrate binding edge of Cu(Z) has a dominantly oxidized Cu(I) and a dominantly reduced Cu(IV). The electronic structure description of Cu(Z) provides a strategy to overcome the reaction barrier of N(2)O reduction at this Cu(I)/Cu(IV) edge by simultaneous two-electron transfer to N(2)O in a bridged binding mode. One electron can be donated directly from Cu(IV) and the other from Cu(II) through the Cu(II)-S-Cu(I) sigma/sigma superexchange pathway. A frontier orbital scheme provides molecular insight into the catalytic mechanism of N(2)O reduction by the Cu(Z) cluster.

}, keywords = {Catalysis, Circular Dichroism, Copper/*chemistry, Crystallography, X-Ray, Electron Spin Resonance Spectroscopy, Histidine/chemistry, Models, Molecular, Oxidoreductases/*chemistry/*metabolism, Spectrum Analysis, Raman/methods, Sulfides/chemistry, Thermodynamics}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=12197752 }, author = {Chen, P. and Cabrito, I. and Moura, J. J. and Moura, I. and Solomon, E. I.} } @article {2339, title = {Heterodimeric nitrate reductase (NapAB) from Cupriavidus necator H16: purification, crystallization and preliminary X-ray analysis}, journal = {Acta Crystallogr Sect F Struct Biol Cryst Commun}, volume = {63}, number = {Pt 6}, year = {2007}, note = {

1744-3091 (Electronic)1744-3091 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {Jun 1}, pages = {516-9}, abstract = {

The periplasmic nitrate reductase from Cupriavidus necator (also known as Ralstonia eutropha) is a heterodimer that is able to reduce nitrate to nitrite. It comprises a 91 kDa catalytic subunit (NapA) and a 17 kDa subunit (NapB) that is involved in electron transfer. The larger subunit contains a molybdenum active site with a bis-molybdopterin guanine dinucleotide cofactor as well as one [4Fe-4S] cluster, while the small subunit is a di-haem c-type cytochrome. Crystals of the oxidized form of this enzyme were obtained using polyethylene glycol 3350 as precipitant. A single crystal grown at the High Throughput Crystallization Laboratory of the EMBL in Grenoble diffracted to beyond 1.5 A at the ESRF (ID14-1), which is the highest resolution reported to date for a nitrate reductase. The unit-cell parameters are a = 142.2, b = 82.4, c = 96.8 A, beta = 100.7 degrees, space group C2, and one heterodimer is present per asymmetric unit.

}, keywords = {Bacterial Proteins/chemistry/*isolation \& purification, Crystallization/methods, Crystallography, X-Ray, Cupriavidus necator/*isolation \& purification, Dimerization, Nitrate Reductase/chemistry/*isolation \& purification, Protein Subunits/chemistry/isolation \& purification}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=17554176 }, author = {Coelho, C. and Gonzalez, P. J. and Trincao, J. and Carvalho, A. L. and Najmudin, S. and Hettman, T. and Dieckman, S. and Moura, J. J. and Moura, I. and Romao, M. J.} } @article {2496, title = {Preliminary crystallographic analysis and further characterization of a dodecaheme cytochrome c from Desulfovibrio desulfuricans ATCC 27774}, journal = {Acta Crystallogr D Biol Crystallogr}, volume = {52}, number = {Pt 6}, year = {1996}, note = {

0907-4449 (Print)0907-4449 (Linking)Journal Article

}, month = {Nov 1}, pages = {1202-8}, abstract = {

Dodecaheme cytochrome c has been purified from Desulfovibrio (D.) desulfuricans ATCC 27774 cells grown under both nitrate and sulfate-respiring conditions. Therefore, it is likely to play a role in the electron-transfer system of both respiratory chains. Its molecular mass (37768 kDa) was determined by electrospray mass spectrometry. Its first 39 amino acids were sequenced and a motif was found between amino acids 32 and 37 that seems to exist in all the cytochromes of the c(3) type from sulfate-reducing bacteria sequenced at present. The midpoint redox potentials of this cytochrome were estimated to be -68, -120, -248 and -310 mV. Electron paramagnetic resonance spectroscopy of the oxidized cytochrome shows several low-spin components with a g(max) spreading from 3.254 to 2.983. Two crystalline forms were obtained by vapour diffusion from a solution containing 2\% PEG 6000 and 0.25-0.75 M acetate buffer pH = 5.5. Both crystals belong to monoclinic space groups: one is P2(1), with a = 61.00, b = 106.19, c = 82.05 A, beta = 103.61 degrees, and the other is C2 with a = 152.17, b = 98.45, c = 89.24 A, beta = 119.18 degrees. Density measurements of the P2(1) crystals suggest that there are two independent molecules in the asymmetric unit. Self-rotation function calculations indicate, in both crystal forms, the presence of a non-crystallographic axis perpendicular to the crystallographic twofold axis. This result and the calculated values for the volume per unit molecular weight of the C2 crystals suggest the presence of two or four molecules in the asymmetric unit.

}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=15299587 }, author = {Coelho, A. V. and Matias, P. M. and Sieker, L. C. and Morais, J. and Carrondo, M. A. and Lampreia, J. and Costa, C. and Moura, J. J. and Moura, I. and Legall, J.} } @article {5881, title = {Induced peroxidase activity of haem containing nitrate reductases revealed by protein film electrochemistry}, journal = {J Electroanal Chem}, volume = {693}, year = {2013}, pages = {105-113}, author = {Coelho, C. and Marangon, J. and Rodrigues, D. and Moura, J. J. G. and Rom{\~a}o, M.J. and Paes de Sousa, P. M. and Correia dos Santos, M. M.} } @article {2273, title = {The crystal structure of Cupriavidus necator nitrate reductase in oxidized and partially reduced states}, journal = {J Mol Biol}, volume = {408}, number = {5}, year = {2011}, note = {

1089-8638 (Electronic)0022-2836 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {May 20}, pages = {932-48}, abstract = {

The periplasmic nitrate reductase (NapAB) from Cupriavidus necator is a heterodimeric protein that belongs to the dimethyl sulfoxide reductase family of mononuclear Mo-containing enzymes and catalyzes the reduction of nitrate to nitrite. The protein comprises a large catalytic subunit (NapA, 91 kDa) containing the molybdenum active site plus one [4Fe-4S] cluster, as well as a small subunit (NapB, 17 kDa), which is a diheme c-type cytochrome involved in electron transfer. Crystals of the oxidized form of the enzyme diffracted beyond 1.5 A at the European Synchrotron Radiation Facility. This is the highest resolution reported to date for a nitrate reductase, providing true atomic details of the protein active center, and this showed further evidence on the molybdenum coordination sphere, corroborating previous data on the related Desulfovibrio desulfuricans NapA. The molybdenum atom is bound to a total of six sulfur atoms, with no oxygen ligands or water molecules in the vicinity. In the present work, we were also able to prepare partially reduced crystals that revealed two alternate conformations of the Mo-coordinating cysteine. This crystal form was obtained by soaking dithionite into crystals grown in the presence of the ionic liquid [C(4)mim]Cl(-). In addition, UV-Vis and EPR spectroscopy studies showed that the periplasmic nitrate reductase from C. necator might work at unexpectedly high redox potentials when compared to all periplasmic nitrate reductases studied to date.

}, keywords = {Amino Acid Sequence, Catalytic Domain, Crystallography, X-Ray, Cupriavidus necator/*enzymology, Cysteine/chemistry, Dithionite/chemistry, Electron Spin Resonance Spectroscopy, Ionic Liquids/chemistry, Molecular Sequence Data, Molybdenum/chemistry, Nitrate Reductase/*chemistry, Periplasm/enzymology, Protein Conformation}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=21419779 }, author = {Coelho, C. and Gonzalez, P. J. and Moura, J. G. and Moura, I. and Trincao, J. and Joao Romao, M.} } @article {2497, title = {Preliminary crystallographic analysis of the oxidized form of a two mono-nuclear iron centres protein from Desulfovibrio desulfuricans ATCC 27774}, journal = {Protein Sci}, volume = {5}, number = {6}, year = {1996}, note = {

0961-8368 (Print)0961-8368 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}tResearch Support, U.S. Gov{\textquoteright}t, P.H.S.

}, month = {Jun}, pages = {1189-91}, abstract = {

Crystals of the fully oxidized form of desulfoferrodoxin were obtained by vapor diffusion from a solution containing 20\% PEG 4000, 0.1 M HEPES buffer, pH 7.5, and 0.2 M CaCl2. Trigonal and/or rectangular prisms could be obtained, depending on the temperature used for the crystal growth. Trigonal prisms belong to the rhombohedral space group R32, with a = 112.5 A and c = 63.2 A; rectangular prisms belong to the monoclinic space group C2, with a = 77.7 A, b = 80.9 A, c = 53.9 A, and beta = 98.1 degrees. The crystallographic asymmetric unit of the rhombohedral crystal form contains one molecule. There are two molecules in the asymmetric unit of the monoclinic form, in agreement with the self-rotation function.

}, keywords = {Crystallization, Crystallography, X-Ray, Desulfovibrio/chemistry/metabolism, Ferredoxins/*chemistry, Oxidation-Reduction}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=8762151 }, author = {Coelho, A. V. and Matias, P. M. and Carrondo, M. A. and Tavares, P and Moura, J. J. and Moura, I. and Fulop, V. and Hajdu, J. and Legall, J.} } @article {2299, title = {Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductase}, journal = {Protein Sci}, volume = {18}, number = {3}, year = {2009}, note = {

1469-896X (Electronic)0961-8368 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {Mar}, pages = {619-28}, abstract = {

Nitric Oxide Reductase (NOR) is an integral membrane protein performing the reduction of NO to N(2)O. NOR is composed of two subunits: the large one (NorB) is a bundle of 12 transmembrane helices (TMH). It contains a b type heme and a binuclear iron site, which is believed to be the catalytic site, comprising a heme b and a non-hemic iron. The small subunit (NorC) harbors a cytochrome c and is attached to the membrane through a unique TMH. With the aim to perform structural and functional studies of NOR, we have immunized dromedaries with NOR and produced several antibody fragments of the heavy chain (VHHs, also known as nanobodies). These fragments have been used to develop a faster NOR purification procedure, to proceed to crystallization assays and to analyze the electron transfer of electron donors. BIAcore experiments have revealed that up to three VHHs can bind concomitantly to NOR with affinities in the nanomolar range. This is the first example of the use of VHHs with an integral membrane protein. Our results indicate that VHHs are able to recognize with high affinity distinct epitopes on this class of proteins, and can be used as versatile and valuable tool for purification, functional study and crystallization of integral membrane proteins.

}, keywords = {Animals, Antibody Affinity, Camelids, New World/*immunology, Crystallography, X-Ray, Epitope Mapping, Humans, Immunoglobulin Fragments/immunology, Immunoglobulin Heavy Chains/immunology, Kinetics, Models, Molecular, Oxidoreductases/*immunology, Peptide Library, Sequence Alignment, Surface Plasmon Resonance}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=19241371 }, author = {Conrath, K. and Pereira, A. S. and Martins, C. E. and Timoteo, C. G. and Tavares, P and Spinelli, S. and Kinne, J. and Flaudrops, C. and Cambillau, C. and Muyldermans, S. and Moura, I. and Moura, J. J. and Tegoni, M. and Desmyter, A.} } @article {2257, title = {Comparative electrochemical study of superoxide reductases}, journal = {Eur Biophys J}, volume = {41}, number = {12}, year = {2012}, month = {Dec 6}, pages = {209-215}, author = {Cordas, C. M. and Raleiras, P. and Auchere, F. and Moura, I. and Moura, J. J.} } @article {2320, title = {Direct electrochemical study of the multiple redox centers of hydrogenase from Desulfovibrio gigas}, journal = {Bioelectrochemistry}, volume = {74}, number = {1}, year = {2008}, note = {

1878-562X (Electronic)1567-5394 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {Nov}, pages = {83-9}, abstract = {

Direct electrochemical response was first time observed for the redox centers of Desulfovibrio gigas [NiFe]-Hase, in non-turnover conditions, by cyclic voltammetry, in solution at glassy carbon electrode. The activation of the enzyme was achieved by reduction with H(2) and by electrochemical control and electrocatalytic activity was observed. The inactivation of the [NiFe]-Hase was also attained through potential control. All electrochemical data was obtained in the absence of enzyme inhibitors. The results are discussed in the context of the proposed mechanism currently accepted for activation/inactivation of [NiFe]-Hases.

}, keywords = {*Electrochemistry, Bacterial Proteins, Desulfovibrio gigas/*enzymology, Electrodes, Enzyme Activation, Hydrogen/chemistry, Hydrogenase/*chemistry/metabolism, Iron, Nickel, Oxidation-Reduction}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=18632311 }, author = {Cordas, C. M. and Moura, I. and Moura, J. J.} } @article {17738, title = {Discovery and characterization of a novel Dyp-type peroxidase from a marine actinobacterium isolated from Trondheim fjord, Norway}, journal = {J Inorg Biochem}, volume = {226}, year = {2022}, pages = {111651}, author = {Cordas, C. M. and G.S. Nguyen and G.V. Val{\'e}rio and M. J{\o}nsson and K. S{\'o}llner and I. Aune and A. Wentzel and Moura, J. J. G.} } @article {3533, title = {Electrochemical behavior of bacterial nitric oxide reductase {\textendash} evidences of low redox potential non-heme FeB give new perspectives on the catalytic mechanism}, journal = {Biochim Biophys Acta}, volume = {1827}, year = {2013}, pages = {233-238}, author = {Cordas, C. M. and Duarte, A. G. and J.J.G .Moura and Moura, I.} } @article {14714, title = {Molybdenum and tungsten enzymes: a brief overview}, journal = {Coord Chem Rev}, volume = {394}, year = {2019}, pages = {53-64}, author = {Cordas, C. M. and Moura, J. J. G.} } @article {2354, title = {Nitric oxide reductase: direct electrochemistry and electrocatalytic activity}, journal = {Chembiochem}, volume = {7}, number = {12}, year = {2006}, note = {

1439-4227 (Print)1439-4227 (Linking)Journal Article

}, month = {Dec}, pages = {1878-81}, abstract = {n/a}, keywords = {Catalysis, Electrochemistry, Oxidation-Reduction, Oxidoreductases/*chemistry}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=17031883 }, author = {Cordas, C. M. and Pereira, A. S. and Martins, C. E. and Timoteo, C. G. and Moura, I. and Moura, J. J. and Tavares, P} } @article {14715, title = {Direct electrochemical reduction of carbon dioxide by a molybdenum-containing formate dehydrogenase}, journal = {J Inorg Biochem}, volume = {196}, year = {2019}, pages = {110694}, url = {https://doi.org/10.1016/j.jinorgbio.2019.110694}, author = {Cordas, C. M. and Campani{\c c}o, M and Baptista, R and Maia, L. and Moura, I. and Moura, J. J. G.} } @article {2338, title = {Simplifying sample handling for protein identification by peptide mass fingerprint using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry}, journal = {Rapid Communications in Mass Spectrometry}, volume = {21}, number = {20}, year = {2007}, note = {

Times Cited: 10

}, month = {2007}, pages = {3269-3278}, abstract = {

An ultrasonic bath, an ultrasonic probe and a sonoreactor were used to speed up the kinetics of the reactions involved in each step of the sample handling for in-gel protein identification by peptide mass fingerprint, PMF, using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The following steps were successfully accelerated using ultrasonic energy: gel washing, protein reduction, and protein alkylation. As a result, a reduction comprising 80\% to 90\% of the total time involved in the classic approach was achieved. In addition the sample handling was also drastically simplified. The number of peptides identified and the protein sequence coverage obtained for the new procedure were comparable to those obtained with the traditional sample treatment for the following protein standards: glycogen phosphorylase b, BSA, ovalbumin, carbonic anhydrase, trypsin inhibitor and alpha-lactalbumin. Finally, as a proof of the procedure, specific proteins were identified from complex protein mixtures obtained from three different sulphate- reducing bacteria: Desulfovibrio, desulfuricans G20, Desulfuvibrio gigas NCIB 9332, and Desulfuvibrio desulfuricans ATCC 27774. Copyright (c) 2007 John Wiley \& Sons, Ltd.

}, isbn = {0951-4198}, url = {http://dx.doi.org/10.1002/rcm.3214}, author = {Cordeiro, Francisco M. and Carreira, Ricardo J. and Rial-Otero, Raquel and Rivas, M. Gabriela and Moura, Isabel and Capelo, Jose-Luis} } @article {2319, title = {Biochemical and spectroscopic characterization of the membrane-bound nitrate reductase from Marinobacter hydrocarbonoclasticus 617}, journal = {J Biol Inorg Chem}, volume = {13}, number = {8}, year = {2008}, note = {

0949-8257 (Print)0949-8257 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {Nov}, pages = {1321-33}, abstract = {

Membrane-bound nitrate reductase from Marinobacter hydrocarbonoclasticus 617 can be solubilized in either of two ways that will ultimately determine the presence or absence of the small (Iota) subunit. The enzyme complex (NarGHI) is composed of three subunits with molecular masses of 130, 65, and 20 kDa. This enzyme contains approximately 14 Fe, 0.8 Mo, and 1.3 molybdopterin guanine dinucleotides per enzyme molecule. Curiously, one heme b and 0.4 heme c per enzyme molecule have been detected. These hemes were potentiometrically characterized by optical spectroscopy at pH 7.6 and two noninteracting species were identified with respective midpoint potentials at Em=+197 mV (heme c) and -4.5 mV (heme b). Variable-temperature (4-120 K) X-band electron paramagnetic resonance (EPR) studies performed on both as-isolated and dithionite-reduced nitrate reductase showed, respectively, an EPR signal characteristic of a [3Fe-4S]+ cluster and overlapping signals associated with at least three types of [4Fe-4S]+ centers. EPR of the as-isolated enzyme shows two distinct pH-dependent Mo(V) signals with hyperfine coupling to a solvent-exchangeable proton. These signals, called "low-pH" and "high-pH," changed to a pH-independent Mo(V) signal upon nitrate or nitrite addition. Nitrate addition to dithionite-reduced samples at pH 6 and 7.6 yields some of the EPR signals described above and a new rhombic signal that has no hyperfine structure. The relationship between the distinct EPR-active Mo(V) species and their plausible structures is discussed on the basis of the structural information available to date for closely related membrane-bound nitrate reductases.

}, keywords = {Alteromonadaceae/*enzymology, Cell Membrane/*metabolism, Electron Spin Resonance Spectroscopy, Electrophoresis, Guanine Nucleotides/analysis/chemistry/metabolism, Heme/chemistry/metabolism, Hydrogen-Ion Concentration, Iron/chemistry/metabolism, Molecular Weight, Molybdenum/chemistry/metabolism, Nitrate Reductase/analysis/*chemistry/*metabolism, Oxidation-Reduction, Potentiometry, Pterins/analysis/chemistry/metabolism, Spectrophotometry, Ultraviolet, Temperature}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=18704520 }, author = {Correia, C. and Besson, S. and Brondino, C. D. and Gonzalez, P. J. and Fauque, G. and Lampreia, J. and Moura, I. and Moura, J. J.} } @article {8444, 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 = {J Biol Inorg Chem}, volume = {20}, year = {2015}, pages = {219-229}, author = {Correia, H. and Marangon, J. and Brondino, C. D. and Moura, J. J. G. and Romao, M. J. and Gonzalez, P. J. and Santos-Silva, T.} } @inbook {3541, title = {Nitrite biosensing using cytochrome c nitrite reductase: Towards a disposable strip electrode}, booktitle = {Biomedical Engineering Systems and Technologies. Series: Communications in Computer and Information Science}, year = {2011}, publisher = {Springer}, organization = {Springer}, author = {Correia, C. and Rodrigues, M. and Silveira, C. M. and Moura, J. J. G. and Ochoteco, E. and Jubete, E. and Almeida, M. G.} } @article {2459, title = {Cross-linking between cytochrome c3 and flavodoxin from Desulfovibrio gigas}, journal = {Biochem Biophys Res Commun}, volume = {256}, number = {2}, year = {1999}, note = {

0006-291X (Print)0006-291X (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {Mar 16}, pages = {367-71}, abstract = {

Tetraheme cytochrome c3 (13 kDa) and flavodoxin (16 kDa), are small electron transfer proteins that have been used to mimic, in vitro, part of the electron-transfer chain that operates between substract electron donors and respiratory electron acceptors partners in Desulfovibrio species (Palma, N., Moura, I., LeGall, J., Van Beeumen, J., Wampler, J., Moura, J. J. G. (1994) Biochemistry 33, 6394-6407). The electron transfer between these two proteins is believed to occur through the formation of a specific complex where electrostatic interaction is the main driving force (Stewart, D., LeGall, J., Moura, I., Moura, J.J.G., Peck, H.D., Xavier, A.V., Weiner, P.K. and Wampler, J.E. (1988) Biochemistry 27, 2444-2450, Stewart, D., LeGall, J., Moura, I., Moura, J.J.G., Peck, H.D., Xavier, A.V., Weiner, P., Wampler, J. (1989) Eur. J. Biochem. 185, 695-700). In order to obtain structural information of the pre-complex, a covalent complex between the two proteins was prepared. A water-soluble carbodiimide [EDC (1-ethyl-3(3 dimethylaminopropyl) carbodiimide hydrochloride] was used for the cross linking reaction. The reaction was optimized varying a wide number of experimental parameters such as ionic strength, protein and cross linker concentration, and utilization of different cross linkers and reaction time between the crosslinker and proteins.

}, keywords = {*Cross-Linking Reagents, Chromatography, High Pressure Liquid, CME-Carbodiimide/analogs \& derivatives, Cytochrome c Group/chemistry/*metabolism, Desulfovibrio/*metabolism, Dose-Response Relationship, Drug, Electron Transport, Ethyldimethylaminopropyl Carbodiimide, Flavodoxin/chemistry/*metabolism, Hydrogen-Ion Concentration, Molecular Weight, Osmolar Concentration, Protein Binding/drug effects, Sodium Chloride/pharmacology, Static Electricity, Succinimides, Time Factors}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=10079190 }, author = {Correia, C. and Monzani, E. and Moura, I. and Lampreia, J. and Moura, J. J.} } @article {2378, title = {Direct electrochemistry of the Desulfovibrio gigas aldehyde oxidoreductase}, journal = {Eur J Biochem}, volume = {271}, number = {7}, year = {2004}, note = {

0014-2956 (Print)0014-2956 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {Apr}, pages = {1329-38}, abstract = {

This work reports on the direct electrochemistry of the Desulfovibrio gigas aldehyde oxidoreductase (DgAOR), a molybdenum enzyme of the xanthine oxidase family that contains three redox-active cofactors: two [2Fe-2S] centers and a molybdopterin cytosine dinucleotide cofactor. The voltammetric behavior of the enzyme was analyzed at gold and carbon (pyrolytic graphite and glassy carbon) electrodes. Two different strategies were used: one with the molecules confined to the electrode surface and a second with DgAOR in solution. In all of the cases studied, electron transfer took place, although different redox reactions were responsible for the voltammetric signal. From a thorough analysis of the voltammetric responses and the structural properties of the molecular surface of DgAOR, the redox reaction at the carbon electrodes could be assigned to the reduction of the more exposed iron cluster, [2Fe-2S] II, whereas reduction of the molybdopterin cofactor occurs at the gold electrode. Voltammetric results in the presence of aldehydes are also reported and discussed.

}, keywords = {Aldehyde Oxidoreductases/*chemistry, Aldehydes, Benzaldehydes/chemistry, Carbon/chemistry, Catalysis, Desulfovibrio gigas/*enzymology, Electrochemistry/*methods, Electron Transport, Gold, Models, Molecular, Oxidation-Reduction, Static Electricity}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=15030483 }, author = {Correia dos Santos, M. M. and Sousa, P. M. and Goncalves, M. L. and Romao, M. J. and Moura, I. and Moura, J. J.} } @article {2458, title = {Electrochemical studies on c-type cytochromes at microelectrodes}, journal = {Journal of Electroanalytical Chemistry}, volume = {464}, number = {1}, year = {1999}, note = {n/a}, pages = {76-84}, type = {doi: 10.1016/S0022-0728(98)00474-4}, abstract = {n/a}, keywords = {Cytochrome c552, Cytochrome c553, Electrochemistry, Horse heart cytochrome c, Macro and microelectrodes}, isbn = {1572-6657}, url = {http://www.sciencedirect.com/science/article/pii/S0022072898004744}, author = {Correia dos Santos, M. M. and Paes de Sousa, P. M. and Sim{\~o}es Gon{\c c}alves, M. L. and Lopes, H. and Moura, I. and Moura, J. J. G.} } @article {2429, title = {Electrochemical studies of rubredoxin from Desulfovibrio vulgaris at modified electrodes}, journal = {Journal of Electroanalytical Chemistry}, volume = {501}, number = {1{\textendash}2}, year = {2001}, note = {n/a}, pages = {173-179}, type = {doi: 10.1016/S0022-0728(00)00521-0}, abstract = {n/a}, keywords = {Modified macro- and micro-electrodes, Rubredoxin, Unmediated electrochemistry}, isbn = {1572-6657}, url = {http://www.sciencedirect.com/science/article/pii/S0022072800005210}, author = {Correia dos Santos, M. M. and Paes de Sousa, P. M. and Sim{\~o}es Gon{\c c}alves, M. L. and Ascenso, C. and Moura, I. and Moura, J. J. G.} } @article {2571, title = {Hexaheme nitrite reductase from Desulfovibrio desulfuricans. Mossbauer and EPR characterization of the heme groups}, journal = {J Biol Chem}, volume = {265}, number = {24}, year = {1990}, note = {

0021-9258 (Print)0021-9258 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}tResearch Support, U.S. Gov{\textquoteright}t, Non-P.H.S.

}, month = {Aug 25}, pages = {14382-8}, abstract = {

Mossbauer and EPR spectroscopy were used to characterize the heme prosthetic groups of the nitrite reductase isolated from Desulfovibrio desulfuricans (ATCC 27774), which is a membrane-bound multiheme cytochrome capable of catalyzing the 6-electron reduction of nitrite to ammonia. At pH 7.6, the as-isolated enzyme exhibited a complex EPR spectrum consisting of a low-spin ferric heme signal at g = 2.96, 2.28, and 1.50 plus several broad resonances indicative of spin-spin interactions among the heme groups. EPR redox titration studies revealed yet another low-spin ferric heme signal at g = 3.2 and 2.14 (the third g value was undetected) and the presence of a high-spin ferric heme. Mossbauer measurements demonstrated further that this enzyme contained six distinct heme groups: one high-spin (S = 5/2) and five low-spin (S = 1/2) ferric hemes. Characteristic hyperfine parameters for all six hemes were obtained through a detailed analysis of the Mossbauer spectra. D. desulfuricans nitrite reductase can be reduced by chemical reductants, such as dithionite or reduced methyl viologen, or by hydrogenase under hydrogen atmosphere. Addition of nitrite to the fully reduced enzyme reoxidized all five low-spin hemes to their ferric states. The high-spin heme, however, was found to complex NO, suggesting that the high-spin heme could be the substrate binding site and that NO could be an intermediate present in an enzyme-bound form.

}, keywords = {Desulfovibrio/*enzymology, Electron Spin Resonance Spectroscopy/methods, Heme/*metabolism, NADH, NADPH Oxidoreductases/*metabolism, Nitrite Reductases/isolation \& purification/*metabolism, Oxidation-Reduction, Spectrum Analysis}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=2167315 }, author = {Costa, C. and Moura, J. J. and Moura, I. and Liu, M. Y. and Peck, H. D., Jr. and Legall, J. and Wang, Y. N. and Huynh, B. H.} } @article {2473, title = {Iron compounds after erythrophagocytosis: chemical characterization and immunomodulatory effects}, journal = {Biochem Biophys Res Commun}, volume = {247}, number = {1}, year = {1998}, note = {

0006-291X (Print)0006-291X (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {Jun 9}, pages = {159-65}, abstract = {

In humans, the lymphomyeloid system has a fundamental role on iron metabolism promoting its recycling due to a continuous removal of effete red blood cells. Additionally, one of the most intriguing aspects of metalloporphyrins in biology is their effect on the immune system. However, the process of erythrocyte catabolism is still poorly understood and needs further research. In the present study, we attempt to investigate the nature and the possible physiologic role of Fe compounds released after erythrophagocytosis during the removal of red blood cells. Monocyte erythrophagocytosis in vitro experiments were done to characterize chemically the Fe compounds present inside the cells and in the culture supernatants. We tested the probable immunomodulatory functions of erythrophagocytosis products over lymphocyte cultures activated in vitro with T mitogens (alpha-CD3). Data obtained from atomic absorption spectroscopy confirmed the presence of Fe in the culture supernatants of monocyte cultures after erythrophagocytosis. Also, high-spin haem complexes derived from erythrocyte catabolism were detected by electron paramagnetic electronic resonance. Finally, in vitro activated lymphocyte proliferation experiments indicate the co-mitogenic properties of monocyte culture supernatants after red blood cells phagocytosis. Thus, the results of the present work provide evidence that culture monocyte supernatants after in vitro erythrophagocytosis contain Fe (III) high-spin haem complexes and show lymphocyte proliferation co-stimulatory properties.

}, keywords = {*Phagocytosis, Adjuvants, Immunologic/blood/chemistry/*physiology, Antibodies, Monoclonal/pharmacology, Antigens, CD3/immunology, Cell-Free System/immunology, Cells, Cultured, Electron Spin Resonance Spectroscopy, Erythrocytes/*chemistry/immunology, Humans, Interleukin-1/analysis/physiology, Iron Compounds/*blood/*chemistry/pharmacology, Lymphocyte Activation/drug effects/immunology, Monocytes/chemistry/immunology, Spectrophotometry, Atomic, Tumor Necrosis Factor-alpha/analysis/physiology}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=9636672 }, author = {Costa, L. M. and Moura, E. M. and Moura, J. J. and de Sousa, M.} } @article {2572, title = {Regulation of the hexaheme nitrite/nitric oxide reductase of Desulfovibrio desulfuricans, Wolinella succinogenes and Escherichia coli. A mass spectrometric study}, journal = {FEBS Lett}, volume = {276}, number = {1-2}, year = {1990}, note = {

0014-5793 (Print)0014-5793 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}tResearch Support, U.S. Gov{\textquoteright}t, Non-P.H.S.

}, month = {Dec 10}, pages = {67-70}, abstract = {

Dissimilatory nitrite reduction, carried out by hexaheme proteins, gives ammonia as the final product. Representatives of this enzyme group from 3 bacterial species can also reduce NO to either ammonia or N2O. The redox regulation of the nitrite/nitric oxide activities is discussed in the context of the denitrifying pathway.

}, keywords = {Ascorbic Acid/metabolism, Bacteroidaceae/*enzymology, Desulfovibrio/*enzymology, Dithionite/metabolism, Escherichia coli/*enzymology, Kinetics, Mass Spectrometry, NADP/metabolism, Nitric Oxide/metabolism, Nitrite Reductases/*metabolism, Oxidoreductases/*metabolism}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=2265715 }, author = {Costa, C. and Macedo, A. and Moura, I. and Moura, J. J. and Legall, J. and Berlier, Y. and Liu, M. Y. and Payne, W. J.} } @article {2481, title = {Formate dehydrogenase from Desulfovibrio desulfuricans ATCC 27774: Isolation and spectroscopic characterization of the active sites (heme, iron-sulfur centers and molybdenum)}, journal = {Journal of Biological Inorganic Chemistry}, volume = {2}, number = {2}, year = {1997}, note = {

Times Cited: 30

}, month = {Apr}, pages = {198-208}, abstract = {

An air-stable formate dehydrogenase, an enzyme that catalyzes the oxidation of formate to CO2, was purified from a sulfate-reducing organism, Desulfovibrio desulfuricans ATCC 27774. The enzyme has a molecular mass of approximately 150 kDa (three different subunits: 88, 29 and 16 kDa) and contains three types of redox-active centers: four c-type hemes, nonheme iron arranged as two [4Fe-4S](2+/1+) centers and a molybdenum-pterin site. Selenium was also chemically detected. The enzyme specific activity is 78 units per mg of protein. Mo(V) EPR signals were observed in the native, reduced and formate-reacted states. EPR signals related to the presence of multiple low-spin hemes were also observed in the oxidized state. Upon reduction, an examination of the EPR data under appropriate conditions distinguishes two types of iron-sulfur centers, an [Fe-S] center I (g(max)=2.050, g(med)=1.947, g(min)=1.896) and an [Fe-S] center II (g(max)=2.071, g(med)=1.926, g(min)=1.865). Mossbauer spectroscopy confirmed the presence of four hemes in the low-spin state. The presence of two [4Fe-4S](2+/1+) centers was confirmed, one of these displaying very small hyperfine coupling constants in the +1 oxidation state. The midpoint redox potentials of the enzyme metal centers were also estimated.

}, isbn = {0949-8257}, url = {http://dx.doi.org/10.1007/s007750050125}, author = {Costa, C. and Teixeira, M. and Legall, J. and Moura, J. J. G. and Moura, I.} } @article {2318, title = {Modelling metallothionein induction in the liver of Sparus aurata exposed to metal-contaminated sediments}, journal = {Ecotoxicology and Environmental Safety}, volume = {71}, number = {1}, year = {2008}, note = {

Times Cited: 10

}, month = {Sep}, pages = {117-124}, abstract = {

Metallothionein (MT) in the liver of gilthead seabreams (Sparus aurata L., 1758) exposed to Sado estuary (Portugal) sediments was quantified to assess the MT induction potential as a biomarker of sediment-based contamination by copper (Cu), cadmium (U), lead (Pb) and arsenic (As). Sediments were collected from two control sites and four sites with different levels of contamination. Sediment Cu, Cd, Pb, As, total organic matter (TOM) and fine fraction (FF) levels were determined. Generalized linear models (GLM) allowed integration of sediment parameters with liver Cu, Cd, Pb, As and MT concentrations. Although sediment metal levels were lower than expected, we relate NIT with liver Cd and also with interactions between liver and sediment Cu and between liver Cu and TOM. We suggest integrating biomarkers and environmental parameters using statistical models such as GLM as a more sensitive and reliable technique for sediment risk assessment than traditional isolated biomarker approaches. (C) 2007 Elsevier Inc. All rights reserved.

}, isbn = {0147-6513}, url = {http://dx.doi.org/10.1016/j.ecoenv.2007.05.012}, author = {Costa, P. M. and Repolho, T. and Caeiro, S. and Diniz, M. E. and Moura, I. and Costa, M. H.} } @article {2495, title = {Redox properties of cytochrome c nitrite reductase from Desulfovibrio desulfuricans ATCC 27774}, journal = {J Biol Chem}, volume = {271}, number = {38}, year = {1996}, note = {

0021-9258 (Print)0021-9258 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}tResearch Support, U.S. Gov{\textquoteright}t, P.H.S.

}, month = {Sep 20}, pages = {23191-6}, abstract = {

The dissimilatory nitrite reductase from Desulfovibrio desulfuricans ATCC 27774 catalyzes the reduction of nitrite to ammonia. Previous spectroscopic investigation revealed that it is a hexaheme cytochrome containing one high spin ferric heme and five low spin ferric hemes in the oxidized enzyme. The current study uses the high resolution of Mossbauer spectroscopy to obtain redox properties of the six heme groups. Correlating the Mossbauer findings with the EPR data reveals the pairwise spin-spin coupling among four of the heme groups. The other two hemes are found to be magnetically isolated. Reduction with dithionite and reaction with CO further indicate that only the high spin heme is capable of binding small exogenous ligands. These results confirm our previous finding that Desulfovibrio desulfuricans nitrite reductase contains six heme groups and that the high spin ferric heme is the substrate and inhibitor binding site.

}, keywords = {*Cytochromes a1, *Cytochromes c1, Carbon Monoxide/chemistry/metabolism, Desulfovibrio/*enzymology, Electron Spin Resonance Spectroscopy, Heme/chemistry/*metabolism, Hydrogenase/metabolism, Nitrate Reductases/chemistry/*metabolism, Oxidation-Reduction, Spectroscopy, Mossbauer, Titrimetry}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=8798514 }, author = {Costa, C. and Moura, J. J. and Moura, I. and Wang, Y. and Huynh, B. H.} } @article {2622, title = {Molybdenum EXAFS of the Desulfovibrio gigas Mo(2Fe-2S) protein--structural similarity to "desulfo" xanthine dehydrogenase}, journal = {J Inorg Biochem}, volume = {20}, number = {4}, year = {1984}, note = {

0162-0134 (Print)0162-0134 (Linking)Comparative StudyJournal ArticleResearch Support, Non-U.S. Gov{\textquoteright}tResearch Support, U.S. Gov{\textquoteright}t, Non-P.H.S.Research Support, U.S. Gov{\textquoteright}t, P.H.S.

}, month = {Apr}, pages = {275-80}, abstract = {

The molybdenum EXAFS of the Mo(2Fe-2S) protein from Desulfovibrio gigas has been examined using fluorescence detection and synchrotron radiation. In the oxidized form the molybdenum environment is found to contain two terminal oxo groups and two long (2.47 A) Mo-S bonds. Evidence was also found for an oxygen or nitrogen donor ligand at 1.90 A. Addition of dithionite to the oxidized enzyme results in loss of a terminal oxo group, perhaps due to protonation. In addition, a 0.1 A contraction in the Mo-S bond lengths is observed. The behavior of both oxidized and dithionite-treated forms is similar to that observed previously with "desulfo" xanthine oxidase.

}, keywords = {*Bacterial Proteins, *Iron-Sulfur Proteins, *Metalloproteins, Desulfovibrio/*analysis, Electron Spin Resonance Spectroscopy, Molybdenum, Protein Conformation, Xanthine Dehydrogenase}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=6325597 }, author = {Cramer, S. P. and Moura, J. J. and Xavier, A. V. and Legall, J.} } @article {2395, 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 = {J Biol Chem}, volume = {278}, number = {19}, year = {2003}, note = {

0021-9258 (Print)0021-9258 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}t

}, month = {May 9}, pages = {17455-65}, abstract = {

The gene encoding cytochrome c nitrite reductase (NrfA) from Desulfovibrio desulfuricans ATCC 27774 was sequenced and the crystal structure of the enzyme was determined to 2.3-A resolution. In comparison with homologous structures, it presents structural differences mainly located at the regions surrounding the putative substrate inlet and product outlet, and includes a well defined second calcium site with octahedral geometry, coordinated to propionates of hemes 3 and 4, and caged by a loop non-existent in the previous structures. The highly negative electrostatic potential in the environment around hemes 3 and 4 suggests that the main role of this calcium ion may not be electrostatic but structural, namely in the stabilization of the conformation of the additional loop that cages it and influences the solvent accessibility of heme 4. The NrfA active site is similar to that of peroxidases with a nearby calcium site at the heme distal side nearly in the same location as occurs in the class II and class III peroxidases. This fact suggests that the calcium ion at the distal side of the active site in the NrfA enzymes may have a similar physiological role to that reported for the peroxidases.

}, keywords = {Amino Acid Sequence, Binding Sites, Calcium/*metabolism, Catalytic Domain/genetics, Cytochrome c Group/analysis/genetics/metabolism, Desulfovibrio/*enzymology, Molecular Sequence Data, Nitrite Reductases/*analysis/genetics/metabolism, Protein Conformation, Sequence Alignment}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=12618432 }, 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. and Moura, I. and Romao, M. J.} } @article {2513, title = {Expression of Desulfovibrio gigas desulforedoxin in Escherichia coli. Purification and characterization of mixed metal isoforms}, journal = {J Biol Chem}, volume = {270}, number = {35}, year = {1995}, note = {

0021-9258 (Print)0021-9258 (Linking)Journal ArticleResearch Support, Non-U.S. Gov{\textquoteright}tResearch Support, U.S. Gov{\textquoteright}t, P.H.S.

}, month = {Sep 1}, pages = {20273-7}, abstract = {

The dsr gene from Desulfovibrio gigas encoding the nonheme iron protein desulforedoxin was cloned using the polymerase chain reaction, expressed in Escherichia coli, and purified to homogeneity. The physical and spectroscopic properties of the recombinant protein resemble those observed for the native protein isolated from D. gigas. These include an alpha 2 tertiary structure, the presence of bound iron, and absorbance maxima at 370 and 506 nm in the UV/visible spectrum due to ligand-to-iron charge transfer bands. Low temperature electron paramagnetic resonance studies confirm the presence of a high-spin ferric ion with g values of 7.7, 5.7, 4.1, and 1.8. Interestingly, E. coli produced two forms of desulforedoxin containing iron. One form was identified as a dimer with the metal-binding sites of both subunits occupied by iron while the second form contained equivalent amounts of iron and zinc and represents a dimer with one subunit occupied by iron and the second with zinc.

}, keywords = {Amino Acid Sequence, Amino Acids/analysis, Bacterial Proteins/biosynthesis, Base Sequence, Binding Sites, Cloning, Molecular, Desulfovibrio/genetics/*metabolism, DNA Primers, Electron Spin Resonance Spectroscopy/methods, Escherichia coli, Genes, Bacterial, Iron-Sulfur Proteins/*biosynthesis/*chemistry/isolation \& purification, Iron/analysis/*metabolism, Mass Spectrometry, Molecular Sequence Data, Polymerase Chain Reaction, Protein Structure, Tertiary, Recombinant Proteins/biosynthesis/chemistry/isolation \& purification, Spectrophotometry}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=PubMed\&dopt=Citation\&list_uids=7657596 }, author = {Czaja, C. and Litwiller, R. and Tomlinson, A. J. and Naylor, S. and Tavares, P and Legall, J. and Moura, J. J. and Moura, I. and Rusnak, F.} } @article {2609, title = {Purification and characterization of three proteins from a halophilic sulfate-reducing bacterium,\<i\>Desulfovibrio salexigens\</i\>}, journal = {Journal of Industrial Microbiology \& Biotechnology}, volume = {1}, number = {3}, year = {1986}, note = {n/a}, pages = {139-147}, publisher = {Springer Berlin / Heidelberg}, abstract = {n/a}, keywords = {Biomedical and Life Sciences}, isbn = {1367-5435}, url = {http://dx.doi.org/10.1007/BF01569265}, author = {Czechowski, M. and Fauque, G. and Galliano, N. and Dimon, B. and Moura, I. and Moura, J. J. G. and Xavier, A. V. and Barato, B. A. S. and Lino, A. R. and Legall, J.} } @article {2608, title = {Resonance Raman spectra of rubredoxin: new assignments and vibrational coupling mechanism from iron-54/iron-56 isotope shifts and variable-wavelength excitation}, journal = {Inorganic Chemistry}, volume = {25}, number = {5}, year = {1986}, note = {

doi: 10.1021/ic00225a022

}, month = {1986/02/01}, pages = {696-700}, publisher = {American Chemical Society}, type = {doi: 10.1021/ic00225a022}, abstract = {n/a}, isbn = {0020-1669}, url = {http://dx.doi.org/10.1021/ic00225a022}, author = {Czernuszewicz, Roman S. and Legall, Jean and Moura, Isabel and Spiro, Thomas G.} }