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71.
Ivan Sazima Cristina Sazima Ronaldo B. Francini-Filho Rodrigo L. Moura 《Environmental Biology of Fishes》2000,59(1):69-77
Gobies of the genus Elacatinus are regarded as the most specialised cleaner fishes in the western tropical Atlantic, yet there are no studies on these cleaners in the southern portion of West Atlantic. We studied the diversity of clients and the daily cleaning activity of the barber goby, Elacatinus figaro, on rocky reefs in southeastern Brazil (23–24°S). A total of 34 fish client species in 16 families were recorded over 484 cleaning events. The most frequent clients were damselfishes, Pomacentridae (37.9% of cleaning events) and grunts, Haemulidae (16.9%). Planktivores were the most frequently attended trophic category, and two species in that category accounted for about a half (44%) of the total cleaning events. Size of clients ranged 4.5–55cm and most individuals were medium-sized (12–30cm); as the barber goby ranged 2–4.5cm, clients were 1.5 to 15 times larger than the cleaner was. Cleaning activity started at dawn and ended shortly before nightfall, the highest frequency of interactions occurring at early morning (nocturnal clients) and mid-afternoon (diurnal clients). By midday the frequency of cleaning events decreased and their duration increased. A total of 109±3 cleaning events and 30±1min of cleaning activity were estimated per cleaning station per day, both figures low when compared to those recorded for cleaner fishes in tropical areas of the western Atlantic and Indo-Pacific. 相似文献
72.
Matthew Kennedy Lian Yu Maria João Lima Carla S. Ascenso Christopher Czaja Isabel Moura Jose J. G. Moura F. Rusnak 《Journal of biological inorganic chemistry》1998,3(6):643-649
Desulforedoxin and the N-terminus of desulfoferrodoxin share a 36 amino acid domain containing a (Cys-S)4 metal binding site. Recombinant forms of desulforedoxin, an N-terminal fragment of desulfoferrodoxin, and two desulforedoxin
mutant proteins were reconstituted with Fe3+, Cd2+, and Zn2+ and relative metal ion affinities assessed by proton titrations. Protons compete with metal for protein ligands, a process
that can be followed by monitoring the optical spectrum of the metal-protein complex as a function of pH. For all polypeptides,
Fe3+ bound with the highest affinity, whereas the affinity of Zn2+ was greater than Cd2+ in desulforedoxin and the N-terminal fragment of desulfoferrodoxin, but this order was reversed in desulforedoxin mutant
proteins. Metal binding in both mutants was significantly impaired. Furthermore, the Fe3+ complex of both mutants underwent a time-dependent bleaching process which coincided with increased reactivity of cysteine
residues to Ellman's reagent and concomitant metal dissociation. It is hypothesized that this results from an autoredox reaction
in which Fe3+ is reduced to Fe2+ with attendant oxidation of ligand thiols.
Received: 17 June 1998 / Accepted: 3 September 1998 相似文献
73.
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75.
M Teixeira I Moura A V Xavier J J Moura J LeGall D V DerVartanian H D Peck B H Huynh 《The Journal of biological chemistry》1989,264(28):16435-16450
The hydrogenase (EC 1.2.2.1) of Desulfovibrio gigas is a complex enzyme containing one nickel center, one [3Fe-4S] and two [4Fe-4S] clusters. Redox intermediates of this enzyme were generated under hydrogen (the natural substrate) using a redox-titration technique and were studied by EPR and M?ssbauer spectroscopy. In the oxidized states, the two [4Fe-4S]2+ clusters exhibit a broad quadrupole doublet with parameters (apparent delta EQ = 1.10 mm/s and delta = 0.35 mm/s) typical for this type of cluster. Upon reduction, the two [4Fe-4S]1+ clusters are spectroscopically distinguishable, allowing the determination of their midpoint redox potentials. The cluster with higher midpoint potential (-290 +/- 20 mV) was labeled Fe-S center I and the other with lower potential (-340 +/- 20 mV), Fe-S center II. Both reduced clusters show atypical magnetic hyperfine coupling constants, suggesting structural differences from the clusters of bacterial ferredoxins. Also, an unusually broad EPR signal, labeled Fe-S signal B', extending from approximately 150 to approximately 450 mT was observed concomitantly with the reduction of the [4Fe-4S] clusters. The following two EPR signals observed at the weak-field region were tentatively attributed to the reduced [3Fe-4S] cluster: (i) a signal with crossover point at g approximately 12, labeled the g = 12 signal, and (ii) a broad signal at the very weak-field region (approximately 3 mT), labeled the Fe-S signal B. The midpoint redox potential associated with the appearance of the g = 12 signal was determined to be -70 +/- 10 mV. At potentials below -250 mV, the g = 12 signal began to decrease in intensity, and simultaneously, the Fe-S signal B appeared. The transformation of the g = 12 signal into the Fe-S signal B was found to parallel the reduction of the two [4Fe-4S] clusters indicating that the [3Fe-4S]o cluster is sensitive to the redox state of the [4Fe-4S] clusters. Detailed redox profiles for the previously reported Ni-signal C and the g = 2.21 signal were obtained in this study, and evidence was found to indicate that these two signals represent two different oxidation states of the enzyme. Finally, the mechanistic implications of our results are discussed. 相似文献
76.
Simone Dell’Acqua Sofia R. Pauleta Isabel Moura José J. G. Moura 《Journal of biological inorganic chemistry》2011,16(2):183-194
This review focuses on the novel CuZ center of nitrous oxide reductase, an important enzyme owing to the environmental significance
of the reaction it catalyzes, reduction of nitrous oxide, and the unusual nature of its catalytic center, named CuZ. The structure
of the CuZ center, the unique tetranuclear copper center found in this enzyme, opened a novel area of research in metallobiochemistry.
In the last decade, there has been progress in defining the structure of the CuZ center, characterizing the mechanism of nitrous
oxide reduction, and identifying intermediates of this reaction. In addition, the determination of the structure of the CuZ
center allowed a structural interpretation of the spectroscopic data, which was supported by theoretical calculations. The
current knowledge of the structure, function, and spectroscopic characterization of the CuZ center is described here. We would
like to stress that although many questions have been answered, the CuZ center remains a scientific challenge, with many hypotheses
still being formed. 相似文献
77.
I Moura A R Lino J J Moura A V Xavier G Fauque H D Peck J LeGall 《Biochemical and biophysical research communications》1986,141(3):1032-1041
Two new low molecular weight proteins with sulfite reductase activity, isolated from Methanosarcina barkeri (DSM 800) and Desulfuromonas acetoxidans (strain 5071), were studied by EPR and optical spectroscopic techniques. Both proteins have visible spectra similar to that of the low-spin sulfite reductase of Desulfovibrio vulgaris strain Hildenborough and no band at 715 nm, characteristic of high-spin Fe3+ complexes in isobacteriochlorins is observed. EPR shows that as isolated the siroheme is in a low-spin ferric state (S = 1/2) with g-values at 2.40, 2.30 and 1.88 for the Methanosarcina barkeri enzyme and g-values at 2.44, 2.33 and 1.81 for the Desulfuromonas acetoxidans enzyme. Chemical analysis shows that both proteins contain one siroheme and one [Fe4S4] center per polypeptidic chain. These results suggest that the low molecular weight, low-spin non-heme iron siroheme proteins represent a new homologous class of sulfite reductases common to anaerobic microorganisms. 相似文献
78.
Raaijmakers H Macieira S Dias JM Teixeira S Bursakov S Huber R Moura JJ Moura I Romão MJ 《Structure (London, England : 1993)》2002,10(9):1261-1272
Desulfovibrio gigas formate dehydrogenase is the first representative of a tungsten-containing enzyme from a mesophile that has been structurally characterized. It is a heterodimer of 110 and 24 kDa subunits. The large subunit, homologous to E. coli FDH-H and to D. desulfuricans nitrate reductase, harbors the W site and one [4Fe-4S] center. No small subunit ortholog containing three [4Fe-4S] clusters has been reported. The structural homology with E. coli FDH-H shows that the essential residues (SeCys158, His159, and Arg407) at the active site are conserved. The active site is accessible via a positively charged tunnel, while product release may be facilitated, for H(+) by buried waters and protonable amino acids and for CO(2) through a hydrophobic channel. 相似文献
79.
de Moura SA de Sousa JM Lima DF Negreiros AN Silva Fde V da Costa LJ 《Gerodontology》2007,24(3):173-176
Objective: The aim of the present study was to analyse the characteristics of salivary production and its composition in individuals with burning mouth syndrome (BMS). Study Design: Salivary flow rate, concentrations of potassium, iron, chloride, thiocyanate, magnesium, calcium, phosphorus, glucose, total protein and urea, as well as the expression profile of salivary proteins were analysed by SDS‐PAGE. Results: The mean salivary flow rate among control patients was lower than that of BMS patients. Chloride, phosphorus and potassium levels were elevated in patients with BMS (p = 0.041, 0.001 and 0.034, respectively). Total salivary protein concentration was reduced in individuals with BMS (p = 0.223). Analysis of the expression of salivary proteins by Coomassie blue SDS‐PAGE revealed a lower expression of low molecular weight proteins in individuals with BMS compared to healthy controls. Conclusions: These results indicate that the identification and characterisation of low molecular weight salivary proteins in BMS may be important in understanding BMS pathogenesis, thus contributing to its diagnosis and treatment. 相似文献
80.
Luís Borlido Leila Moura Ana M. Azevedo Ana C. A. Roque Dr. Maria R. Aires-Barros Dr. José Paulo S. Farinha 《Biotechnology journal》2013,8(6):709-717
Monoclonal antibodies (mAbs) are important therapeutic proteins. One of the challenges facing large-scale production of monoclonal antibodies is the capacity bottleneck in downstream processing, which can be circumvented by using magnetic stimuli-responsive polymer nanoparticles. In this work, stimuli-responsive magnetic particles composed of a magnetic poly(methyl methacrylate) core with a poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAM-co-AA)) shell cross-linked with N, N'-methylenebisacrylamide were prepared by miniemulsion polymerization. The particles were shown to have an average hydrodynamic diameter of 317 nm at 18°C, which decreased to 277 nm at 41°C due to the collapse of the thermo-responsive shell. The particles were superparamagnetic in behavior and exhibited a saturation magnetization of 12.6 emu/g. Subsequently, we evaluated the potential of these negatively charged stimuli-responsive magnetic particles in the purification of a monoclonal antibody from a diafiltered CHO cell culture supernatant by cation exchange. The adsorption of antibodies onto P(NIPAM-co-AA)-coated nanoparticles was highly selective and allowed for the recovery of approximately 94% of the mAb. Different elution strategies were employed providing highly pure mAb fractions with host cell protein (HCP) removal greater than 98%. By exploring the stimuli-responsive properties of the particles, shorter magnetic separation times were possible without significant differences in product yield and purity. 相似文献