Two-dimensional 1H NMR studies on Desulfovibrio gigas ferredoxins. Assignment of the iron-sulfur cluster cysteinyl ligand protons,
Macedo, Anjos L., Palma Nuno P., Moura Isabel, Legall Jean, Wray Victor, and Moura José J. G.
, Magnetic Resonance in Chemistry, Volume 31, Number 13, p.S59-S67, (1993)
Abstract1D and 2D 1H NMR studies are reported on the oxidized and reduced [4Fe-4S] cluster of Desulfovibrio gigas ferredoxin I (Fdl). Several low-field contact shifted resonances (fast relaxing) are assigned to β-CH2 and α-CH coordinated cysteinyl residues. NOESY patterns (supported by 1D NOE experiments) resolves four pairs of geminal β-CH2 protons at low-field. The cluster ligands are assigned non-specifically to Cys8, Cys11, Cys14 and Cys50, based on the X-ray structural analysis available for the oligomeric form, FdII, that contains a single [3Fe-4S] cluster. It was indicated in this case that Cys11 is not bound to the trinuclear cluster but is tilted towards the solvent. The presence of four pairs of geminal β-CH2 protons for FdI unambiguously proves the occupancy of the fourth site of the [3Fe-4S] complex and implies the coordination of the Cys11 at the cluster. Analysis of the oxidized form of FdII, using the same methodology as described for FdI, supports the presence of three cysteinyl ligands in the [3Fe-4S] core. Further, the combined use of the X-ray coordinates enables the specific assignment of the three cysteinyl ligands of the cluster, extending a previous assignment of Cys50. In addition, very broad resonances were detected for the reduced form of FdII in the low-field region around 200 ppm and in the high field region around −80 ppm.
Two azurins with unusual redox and spectroscopic properties isolated from the Pseudomonas chlororaphis strains DSM 50083(T) and DSM 50135,
Pinho, D., Besson S., Brondino C. D., Pereira E., de Castro B., and Moura I.
, Journal of Inorganic Biochemistry, Feb, Volume 98, Number 2, p.276-286, (2004)
AbstractTwo azurins (Az624 and Az626) were isolated from the soluble extract of two strains of Pseudomonas chlororaphis, DSM 50083(T) and DSM 50135, respectively, grown under microaerobic conditions with nitrate as final electron acceptor. The azurins, purified to electrophoretic homogeneity in three chromatographic steps, exhibit several peculiar properties. They have high reduction potentials and lower pI than most azurins described in the literature. As previously observed for Pseudomonas aeruginosa azurin, their reduction potentials are pH-dependent, but the pK values of their oxidized forms are lower, which suggests that deeper structural changes are associated with the oxidation process of these novel azurins. A hitherto undescribed pH-dependence of the diffusion coefficient was observed in Az624, that could be caused either by conformational changes, or by the formation of supramolecular aggregates associated with a protonation process. Both azurins exhibit axial X-band electron paramagnetic resonance spectra in frozen solution showing a typical hyperfine with the copper nucleus (I = 3/2) and a well-resolved superhyperfine structure with two equivalent N-14 nucleus (I = 1), which is not usually observed for azurins from other species. (C) 2003 Elsevier Inc. All rights reserved.
Tungsten-containing formate dehydrogenase from Desulfovibrio gigas: metal identification and preliminary structural data by multi-wavelength crystallography,
Raaijmakers, H., Teixeira S., Dias J. M., Almendra M. J., Brondino C. D., Moura I., Moura J. J., and Romao M. J.
, J Biol Inorg Chem, Apr, Volume 6, Number 4, p.398-404, (2001)
AbstractThe tungsten-containing formate dehydrogenase (W-FDH) isolated from Desulfovibrio gigas has been crystallized in space group P2(1), with cell parameters a = 73.8 A, b = 111.3 A, c = 156.6 A and beta = 93.7 degrees. These crystals diffract to beyond 2.0 A on a synchrotron radiation source. W-FDH is a heterodimer (92 kDa and 29 kDa subunits) and two W-FDH molecules are present in the asymmetric unit. Although a molecular replacement solution was found using the periplasmic nitrate reductase as a search model, additional phasing information was needed. A multiple-wavelength anomalous dispersion (MAD) dataset was collected at the W- and Fe-edges, at four different wavelengths. Anomalous and dispersive difference data allowed us to unambiguously identify the metal atoms bound to W-FDH as one W atom with a Se-cysteine ligand as well as one [4Fe-4S] cluster in the 92 kDa subunit, and three additional [4Fe-4S] centers in the smaller 29 kDa subunit. The D. gigas W-FDH was previously characterized based on metal analysis and spectroscopic data. One W atom was predicted to be bound to two molybdopterin guanine dinucleotide (MGD) pterin cofactors and two [4Fe-4S] centers were proposed to be present. The crystallographic data now reported reveal a selenium atom (as a Se-cysteine) coordinating to the W site, as well as two extra [4Fe-4S] clusters not anticipated before. The EPR data were re-evaluated in the light of these new results.
Total synthesis of a simple metalloprotein-desulforedoxin,
Tavares, P., Wunderlich J. K., Lloyd S. G., Legall J., Moura J. J., and Moura I.
, Biochem Biophys Res Commun, Mar 17, Volume 208, Number 2, p.680-7, (1995)
AbstractDesulforedoxin is a protein purified from cellular extracts of Desulfovibrio gigas. It is a small (7.9 kDa) dimeric protein that contains a distorted rubredoxin like center (one single iron coordinated by four cysteinyl residues). Due to the simplicity of the polypeptide chain and of the iron center, an attempt was made to chemically produce this protein. A 36 amino acid polypeptide chain was synthesized based on the known sequence of native Desulforedoxin. The iron center was then reconstituted and the biochemical and spectroscopic characteristics of this synthetic protein were investigated. The final product has an equal sequence to the protein purified from D. gigas. The synthetic and natural Dx are very similar, in terms redox potential and spectroscopic properties (UV-Visible, EPR, Mossbauer).
Total lead and its stable isotopes in the digestive gland of Octopus vulgaris as a fingerprint,
Raimundo, J., Vale C., Caetano M., Cesario R., and Moura I.
, Aquatic Biology, 2009, Volume 6, Number 1-3, p.25-30, (2009)
AbstractWe hypothesised that the isotopic signature of Pb in the digestive gland of the common octopus reflects the organisms' sources of Pb, and investigated whether isotopic Pb ratios are useful in characterising octopus populations. A total of 47 Octopus vulgaris individuals were captured between November 2005 and September 2006 in 2 areas of the Portuguese coast, near Matosinhos (Area A; NW coast) and Olhao (Area B; south coast), and digestive glands were analysed for total Pb and its stable isotopes. The same determinations were performed in 22 samples of surface sediments from the 2 areas. Pb concentrations in the digestive gland of specimens from Area B (2.8 to 13.0 mu g g(-1)) exceeded the values found in Area A (1.3 to 8.3 mu g g(-1)). A similar pattern was found for the isotopic Pb ratios: (206)Pb/(207)Pb was 1.173 to 1.185 for Area A and 1.165 to 1.172 for B; (206)Pb/(208)Pb was 0.476 to 0.487 for Area A and 0.318 to 0.483 for B. The different signatures of the digestive glands are in line with those observed in the surface sediments of the 2 coastal areas (e.g. (206)Pb/(207)Pb was 1.179 to 1.207 for Area A and 1.171 to 1.181 for B). However, the isotopic Pb signature of octopus was less radiogenic than that of sediments. Because octopus has a short life span (up to 24 mo) the signature reflects recent sources of Pb that have a less radiogenic signature. The Pb signature of surface sediments tends to integrate the record of the previous few years or decades, due to the frequent resuspension of the upper layer of coastal sediments. The mixing of sediments deposited during those periods results in higher isotopic Pb ratios (more radiogenic). The consistent differences between the 2 areas, in sediments and octopus, points towards the isotopic Pb signature as a possible useful tool to distinguish octopus populations.
Topography of human cytochrome b5/cytochrome b5 reductase interacting domain and redox alterations upon complex formation,
Samhan-Arias, A. K., Almeida R. M., Ramos S., Cordas C. M., Moura I., Gutierrez-Merino C., and Moura J. J. G.
, Biochim Biophys Acta, Volume 1859, p.78-87, (2018)
Three-iron clusters in iron--sulfur proteins: An EPR study of the exchange interactions,
Gayda, Jean-Pierre, Bertrand Patrick, Theodule Francois-Xavier, and Moura Jose J. G.
, The Journal of Chemical Physics, Volume 77, Number 7, p.3387-3391, (1982)
Abstractn/a