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in vitro biosynthesis of poly(3-hydroxybutyric acid) by using purified poly(hydroxyalkanoic acid) synthase of chromatium vinosum.purified recombinant poly(hydroxyalkanoic acid) (pha) synthase from chromatium vinosum (phaeccv) was used to examine in vitro the specific synthase activity, turnover of r-(-)-3-hydroxybutyryl coenzyme a (3hb-coa) and poly(3-hydroxybutyric acid) formation under various conditions. the 3hb-coa consumption was terminated by a reaction-dependent inactivation of the pha synthase. salts (mgcl2, cacl2, nacl), proteins (bovine serum albumin, lysozyme, phasine) or detergent (tween 20) increased the 3hb- ...19989581289
the two [4fe-4s] clusters in chromatium vinosum ferredoxin have largely different reduction potentials. structural origin and functional consequences.the 2[4fe-4s] ferredoxin from chromatium vinosum arises as one prominent member of a recently defined family of proteins found in very diverse bacteria. the potentiometric circular dichroism titrations of the protein and of several molecular variants generated by site-directed mutagenesis have established that the reduction potentials of the two clusters differ widely by almost 200 mv. this large difference has been confirmed by electrochemical methods, and each redox transition has been assigne ...19989624123
sulfide oxidation in the phototrophic sulfur bacterium chromatium vinosum.sulfide oxidation in the phototrophic purple sulfur bacterium chromatium vinosum d (dsmz 180(t)) was studied by insertional inactivation of the fccab genes, which encode flavocytochrome c, a protein that exhibits sulfide dehydrogenase activity in vitro. flavocytochrome c is located in the periplasmic space as shown by a phoa fusion to the signal peptide of the hemoprotein subunit. the genotype of the flavocytochrome-c-deficient chr. vinosum strain fd1 was verified by southern hybridization and p ...19989639604
evidence against the double-arginine motif as the only determinant for protein translocation by a novel sec-independent pathway in escherichia coli.proteins which are synthesized with a signal peptide containing a 'double-arginine' motif may be translocated across the bacterial cytoplasmic membrane by a mechanism that is different from the known sec and signal recognition particle pathways. the function of the double-arginine motif as a determinant for this novel pathway was studied by expressions of gene constructs coding for the high potential iron-sulfur protein (hipip) from chromatium vinosum d in escherichia coli. when the protein was ...19989682482
synechocystis sp. pcc6803 possesses a two-component polyhydroxyalkanoic acid synthase similar to that of anoxygenic purple sulfur bacteria.during cultivation under storage conditions with bg11 medium containing acetate as a carbon source, synechocystis sp. pcc6803 accumulated poly(3-hydroxybutyrate) up to 10% (w/w) of the cell dry weight. our analysis of the complete synechocystis sp. pcc6803 genome sequence, which had recently become available, revealed that not only the open reading frame slr1830 (which was designated as phac) but also the open reading frame slr1829, which is located colinear and upstream of phac, most probably r ...19989683655
sirohaem sulfite reductase and other proteins encoded by genes at the dsr locus of chromatium vinosum are involved in the oxidation of intracellular sulfur.the sequence of the dsr gene region of the phototrophic sulfur bacterium chromatium vinosum d (dsmz 180) was determined to clarify the in vivo role of 'reverse' sirohaem sulfite reductase. the dsrab genes encoding dissimilatory sulfite reductase are part of a gene cluster, dsrabefhcmk, that encodes four small, soluble proteins (dsre, dsrf, dsrh and dsrc), a transmembrane protein (dsrm) with similarity to haem-b-binding polypeptides and a soluble protein (dsrk) resembling [4fe-4s]-cluster-contain ...19989695921
crystal structure and possible dimerization of the high-potential iron-sulfur protein from chromatium purpuratum.the crystal structure of the high-potential iron-sulfur protein (hipip) isolated from chromatium purpuratum is reported at 2.7 a resolution. the three hipip molecules in the asymmetric unit of the crystals form one and one-half dimers. two molecules are related by a noncrystallographic symmetry rotation of approximately 175 degrees with negligible translation along the dyad axis. the third molecule in the asymmetric unit also forms a dimer with a second hipip molecule across the crystallographic ...19989760225
purification and characterization of chromatium vinosum groel and groes proteins overexpressed in escherichia coli cells lacking the endogenous groesl operon.using an escherichia coli strain (rf101) in which the endogenous chromosomal groesl operon was removed, we overexpressed the groel and groes chaperonins cloned from the photosynthetic bacterium chromatium vinosum. the identities of these proteins were confirmed by immunological and n-terminal sequence analyses. the native molecular masses of groel and groes, as determined by size-exclusion chromatography, were 830 and 74 kda, respectively. this suggests a tetradecameric structure for groel and a ...19989790891
acclimation of the photosynthetic response of chromatium vinosum to light-limiting conditions.the photosynthetic response of the purple sulfur bacterium chromatium vinosum dsm 185 to different degrees of illumination was analyzed. the microorganism was grown in continuous culture, and samples were taken from the effluent of the culture and incubated at different irradiances to determine the specific rate of sulfur oxidation as a measure of the photosynthetic activity of the organism. the activities obtained were plotted as a function of the specific rate of light uptake, and for each set ...19989799283
utilization of reducing power in light-limited cultures of chromatium vinosum dsm 185.this study describes how the phototrophic organism chromatium vinosum, when grown under different degrees of light limitation, distributes the reducing power initially present in the medium as hydrogen sulfide. under all the conditions of illumination tested, sulfur was the major store of reducing power. glycogen, which was virtually absent under light limitation, accounted for 31.6% of the stored reducing power at saturating irradiances. analysis of the electron budget showed that under light-l ...19989799284
basis for monomer stabilization in rhodopseudomonas palustris cytochrome c' derived from the crystal structure.the crystal structure of an unusual monomeric cytochrome c' from rhodopseudomonas palustris (rpcp) has been determined at 2.3 a resolution. rpcp has the four-helix (helices a, b, c and d) bundle structure similar to dimeric cytochromes c'. however the amino acid composition of the surface of helices a and b in rpcp is remarkably different from that of the dimeric cytochromes c'. this surface forms the dimer interface in the latter proteins. rpcp has seven charged residues on this surface contrar ...19989826513
phylogenetic relationships among the chromatiaceae, their taxonomic reclassification and description of the new genera allochromatium, halochromatium, isochromatium, marichromatium, thiococcus, thiohalocapsa and thermochromatium.sequences of the 16s rdna from all available type strains of chromatium species have been determined and were compared to those of other chromatiaceae, a few selected ectothiorhodospiraceae and escherichia coli. the clear separation of ectothiorhodospiraceae and chromatiaceae is confirmed. most significantly the sequence comparison revealed a genetic divergence between chromatium species originated from freshwater sources and those of truly marine and halophilic nature. major phylogenetic branch ...19989828415
in vitro synthesis of poly(3-hydroxybutyric acid) by using an enzymatic coenzyme a recycling system.purified recombinant poly(hydroxyalkanoic acid), pha, synthase from chromatium vinosum was used to examine in vitro poly(3-hydroxybutyric acid) (p(3hb)) formation. in combination with purified propionyl-coenzyme a transferase of clostridium propionicum a two-enzyme in vitro p(3hb) biosynthesis system was established which allowed the synthesis of p(3hb) from free d-(-)-3-hydroxybutyric acid as substrate. the coenzyme a residue for the activation of this hydroxyacid was provided by acetyl-coenzym ...19989835044
effect of modification of histidine residues on organization of the pigment--protein complexes of chromatium minutissimumthe effect of diethyl pyrocarbonate on chromatophores and isolated pigment--protein complexes of chromatium minutissimum was studied. it is shown that modification of histidine residues results in the destruction of the core antenna lhi (b880) and in a spectral shift from 850 to 830 nm in the peripheral antenna lhii (b800-850). in the purple sulfur bacterium chromatium minutissimum the pigment--protein complexes b800-b850 (peripheral antenna, lhii) and b880 (core antenna, lhi) collect and transm ...19989864456
pha synthase from chromatium vinosum: cysteine 149 is involved in covalent catalysis.polyhydroxyalkanoate synthase (pha) from chromatium vinosum catalyzes the conversion of 3-hydroxybutyryl-coa (hb-coa) to polyhydroxybutyrate (phb) and coa. the synthase is composed of a approximately 1:1 mixture of two subunits, phac and phae. size-exclusion chromatography indicates that in solution phac and phae exist as large molecular weight aggregates. the holo-enzyme, phaec, has a specific activity of 150 units/mg. each subunit was cloned, expressed, and purified as a (his)6-tagged construc ...19999888824
carbon monoxide and cyanide as intrinsic ligands to iron in the active site of [nife]-hydrogenases. nife(cn)2co, biology's way to activate h2.infrared-spectroscopic studies on the [nife]-hydrogenase of chromatium vinosum-enriched in 15n or 13c, as well as chemical analyses, show that this enzyme contains three non-exchangeable, intrinsic, diatomic molecules as ligands to the active site, one carbon monoxide molecule and two cyanide groups. the results form an explanation for the three non-protein ligands to iron detected in the crystal structure of the desulfovibrio gigas hydrogenase (volbeda, a., garcin, e., piras, c., de lacey, a. i ...19999920874
taxon-specific content of oligonucleotide triplets in 16s rrnas of anoxygenic phototrophic and nitrifying bacteria.theoretical evaluation of the content of oligonucleotide triplets aaa, ccc, and uau in 16s rrnas of anoxygenic phototrophic bacteria (genera chlorobium; chloroflexus; chromatium: rhodopseudomonas) and nitrifying bacteria (genera nitrosococcus, nitrosomonas, nitrosolobus, nitrosovibrio, nitrospira, nitrospina, nitrobacter) showed that the number of the aaa, ccc or uau triplets in 16s rrnas specifically corresponds to the genus and species of bacteria. the ratio of aaa and ccc triplet numbers in t ...199910049622
diversity of dissimilatory bisulfite reductase genes of bacteria associated with the deep-sea hydrothermal vent polychaete annelid alvinella pompejana.a unique community of bacteria colonizes the dorsal integument of the polychaete annelid alvinella pompejana, which inhabits the high-temperature environments of active deep-sea hydrothermal vents along the east pacific rise. the composition of this bacterial community was characterized in previous studies by using a 16s rrna gene clone library and in situ hybridization with oligonucleotide probes. in the present study, a pair of pcr primers (p94-f and p93-r) were used to amplify a segment of th ...199910049872
in situ analysis of phototrophic sulfur bacteria in the chemocline of meromictic lake cadagno (switzerland).comparative sequence analysis of a 16s rrna gene clone library from the chemocline of the meromictic lake cadagno (switzerland) revealed the presence of a diverse number of phototrophic sulfur bacteria. sequences resembled those of rrna of type strains chromatium okenii dsm169 and amoebobacter purpureus dsm4197, as well as those of four bacteria forming a tight cluster with a. purpureus dsm4197 and lamprocystis roseopersicina dsm229. in situ hybridization with fluorescent (cy3 labeled) oligonucl ...199910049902
pre-steady-state kinetics of the reactions of [nife]-hydrogenase from chromatium vinosum with h2 and co.results are presented of the first rapid-mixing/rapid-freezing studies with a [nife]-hydrogenase. the enzyme from chromatium vinosum was used. in particular the reactions of active enzyme with h2 and co were monitored. the conversion from fully reduced, active hydrogenase (nia-sr state) to the nia-c* state was completed in less than 8 ms, a rate consistent with the h2-evolution activity of the enzyme. the reaction of co with fully reduced enzyme was followed from 8 to 200 ms. the nia-sr state di ...199910092843
phylogeny and diversity of achromatium oxaliferum.achromatium oxaliferum was first described in 1893 by schewiakoff as an unusually large bacterium living in freshwater sediments. up to now no pure culture is available. physical enrichments of achromatia collected from the acidic lake fuchskuhle, which houses a peculiar, smaller variety, and the neutral lake stechlin were investigated by the cultivation-independent rrna approach. pcr in combination with cloning and sequencing was used for the retrieval of 24 partial and 4 nearly full-length 16s ...199910188276
structural and dynamical properties of a partially unfolded fe4s4 protein: role of the cofactor in protein folding.heteronuclear multidimensional nmr spectroscopy was used to investigate in detail the structural and dynamical properties of a partially unfolded intermediate of the reduced high-potential iron-sulfur protein (hipip) from chromatium vinosum present in 4 m guanidinium chloride solution. after an extensive assignment of 15n and 1h resonances, noe data, proton longitudinal relaxation times, and 3jhnhalpha coupling constants as well as 15n relaxation parameters (t1, t2, t1rho, and 1h-15n noe) were o ...199910200154
kinetics of photoacclimation in cultures of chromatium vinosum dsm 185 during shifts in light irradiance.continuous cultures of chromatium vinosum dsm 185 were shifted from a high to a low irradiance (67 to 4 microe m(-2) s(-1)) and vice versa (4 to 67 microe m(-2) s(-1)). the kinetics of photoacclimation of the cultures were analysed during these transitions until steady state was reached. when irradiance was shifted from 4 to 67 microe m(-2) s(-1), bacteriochlorophyll synthesis halted for 4 h. during this period, pigments were progressively diluted in the newly formed biomass, resulting in a lowe ...199910220162
unusual nmr, epr, and mössbauer properties of chromatium vinosum 2[4fe-4s] ferredoxin.the ferredoxin from chromatium vinosum (cvfd) exhibits sequence and structure peculiarities. its two fe4s4(scys)4 clusters have unusually low potential transitions that have been unambiguously assigned here through nmr, epr, and mössbauer spectroscopy in combination with site-directed mutagenesis. the [4fe-4s]2+/1+ cluster (cluster ii) whose coordination sphere includes a two-turn loop between cysteines 40 and 49 was reduced by dithionite with an e degrees ' of -460 mv. its s = 1/2 epr signal wa ...199910320364
mad structure of pseudomonas nautica dimeric cytochrome c552 mimicks the c4 dihemic cytochrome domain association.the monohemic cytochrome c552from pseudomonas nautica (c552-pn) is thought to be the electron donor to cytochrome cd1, the so-called nitrite reductase (nir). it shows as high levels of activity and affinity for the p. nautica nir (nir-pn), as the pseudomonas aeruginosa enzyme (nir-pa). since cytochrome c552is by far the most abundant electron carrier in the periplasm, it is probably involved in numerous other reactions. its sequence is related to that of the c type cytochromes, but resembles tha ...199910369779
uphill energy transfer in lh2-containing purple bacteria at room temperatureuphill energy transfer in the lh2-containing purple bacteria rhodopseudomonas acidophila, rhodopseudomonas palustris, rhodobacter sphaeroides, chromatium vinosum and chromatium purpuratum was studied by stationary fluorescence spectroscopy at room temperature upon selective excitation of the b800 pigments of lh2 and the b880 pigments of lh1 at 803 nm and 900 nm, respectively. the resulting fluorescence spectra differed significantly at wavelengths shorter than the fluorescence maximum but agreed ...199910393258
catalytic electron transport in chromatium vinosum [nife]-hydrogenase: application of voltammetry in detecting redox-active centers and establishing that hydrogen oxidation is very fast even at potentials close to the reversible h+/h2 value.the nickel-iron hydrogenase from chromatium vinosum adsorbs at a pyrolytic graphite edge-plane (pge) electrode and catalyzes rapid interconversion of h(+)((aq)) and h(2) at potentials expected for the half-cell reaction 2h(+) right arrow over left arrow h(2), i.e., without the need for overpotentials. the voltammetry mirrors characteristics determined by conventional methods, while affording the capabilities for exquisite control and measurement of potential-dependent activities and substrate-pr ...199910413472
investigation of the role of a surface patch in the self-association of chromatium vinosum high potential iron-sulfur protein.the role of a flattened, relatively hydrophobic surface patch in the self-association of chromatium vinosum hipip was assessed by substituting phenylalanine 48 with lysine. the reduction potential of the f48k variant was 26 mv higher than that of the wild-type (wt) recombinant (rc) hipip, consistent with the introduction of a positive charge close to the cluster. nuclear magnetic resonance spectroscopy (nmr) revealed that the electronic structure of the oxidized cluster in these two proteins is ...199910446369
amino acid sequences of two high-potential iron-sulfur proteins (hipips) from the moderately halophilic purple phototrophic bacterium, rhodospirillum salinarum.the amino acid sequences of two very different high-potential iron-sulfur protein (hipip) isozymes have been determined from the moderately halophilic purple phototrophic bacterium, rhodospirillum salinarum. iso-1 hipip, which is monomeric and contains 57 amino acid residues, is most similar to the thiobacillus ferrooxidans iron-oxidizing enzyme (45% identity and a 6-residue deletion). on the other hand, iso-2 hipip, which is isolated as an oligomer, contains a peptide chain with 54 amino acid r ...199910462450
ab initio solution and refinement of two high-potential iron protein structures at atomic resolution.the crystal structure of the reduced high-potential iron protein (hipip) from chromatium vinosum has been redetermined in a new orthorhombic crystal modification, and the structure of its h42q mutant has been determined in orthorhombic (h42q-1) and cubic (h42q-2) modifications. the first two were solved by ab initio direct methods using data collected to atomic resolution (1.20 and 0. 93 a, respectively). the recombinant wild type (rc-wt) with two hipip molecules in the asymmetric unit has 1264 ...199910531472
an unusual arrangement of pur and lpx genes in the photosynthetic purple sulfur bacterium allochromatium vinosum.the nucleotide sequence of a 1634 bp dna fragment from the photosynthetic purple sulfur bacterium allochromatium vinosum contains one complete and two partial open reading frames. sequence comparisons to genes from other organisms suggest that this a. vinosum dna fragment contains, starting from the 5' end, the following: (1) 234 bp at the 3' end of the a. vinosum purh gene, coding for 78 amino acids at the c-terminus of the bi-functional 5'-phosphoribosyl-5-aminoimidazole-4-carboxamide formyltr ...199910532315
orientation-selected endor of the active center in chromatium vinosum [nife] hydrogenase in the oxidized "ready" state.electron nuclear double resonance (endor) was applied to study the active site of the oxidized "ready" state, ni(r), in the [nife] hydrogenase of chromatium vinosum. the magnetic field dependence of the epr was used to select specific subsets of molecules contributing to the endor response by stepping through the epr envelope. three hyperfine couplings could be clearly followed over the complete field range. two protons, h1 and h2, display a very similar large isotropic coupling of 12.5 and 12.6 ...199910555572
certain species of the proteobacteria possess unusual bacteriochlorophyll a environments in their light-harvesting proteins.in this work, we have examined, using fourier-transform raman (ft-r) spectroscopy, the bacteriochlorophyll a (bchl a) binding sites in light-harvesting (lh) antennae from different species of the proteobacteria that exhibit unusal absorption properties. while the lh1 complexes from erythromicrobium (e.) ramosum (rc-b871) and rhodospirillum centenum (b875) present classic ft-r spectra in the carbonyl high-frequency region, we show that in the blue-shifted lh1 complex, absorbing at 856 nm, from ro ...199910604286
experimental evidence for the role of buried polar groups in determining the reduction potential of metalloproteins: the s79p variant of chromatium vinosum hipip.the amide group between residues 78 and 79 of chromatium vinosum high-potential iron-sulfur protein (hipip) is in close proximity to the fe4s4 cluster of this protein and interacts via a hydrogen bond with s gamma of cys77, one of the cluster ligands. the reduction potential of the s79p variant was 104 +/- 3 mv lower than that of the recombinant wild-type (rcwt) hipip (5 mm phosphate, 100 mm nacl, ph 7, 293 k), principally due to a decrease in the enthalpic term which favors the reduction of the ...199910631600
lipases provide a new mechanistic model for polyhydroxybutyrate (phb) synthases: characterization of the functional residues in chromatium vinosum phb synthase.polyhydroxybutyrate (phb) synthases catalyze the conversion of beta-hydroxybutyryl coenzyme a (hbcoa) to phb. these enzymes require an active site cysteine nucleophile for covalent catalysis. a protein blastp search using the class iii chromatium vinosum synthase sequence reveals high homology to prokaryotic lipases whose crystal structures are known. the homology is very convincing in the alpha-beta-elbow (with the active site nucleophile)-alpha-beta structure, residues 131-175 of the synthase. ...200010747780
unusual spectroscopic and electrochemical properties of the 2[4fe-4s] ferredoxin of thauera aromatica.a reduced ferredoxin serves as the natural electron donor for key enzymes of the anaerobic aromatic metabolism in the denitrifying bacterium thauera aromatica. it contains two [4fe-4s] clusters and belongs to the chromatium vinosum type of ferredoxins (cvfd) which differ from the "clostridial" type by a six-amino acid insertion between two successive cysteines and a c-terminal alpha-helical amino acid extension. the electrochemical and electron paramagnetic resonance (epr) spectroscopic properti ...200010769152
crystallization and preliminary crystallographic analysis of the high-potential iron-sulfur protein from thermochromatium tepidum.the high-potential iron-sulfur protein (hipip) is an electron carrier between the photosynthetic reaction centre and the cytochrome bc(1) complex in the electron-transfer chain of photosynthesis. the purified hipip from thermochromatium tepidum (formerly chromatium tepidum) was crystallized in a solution of 1.4 m ammonium sulfate and 0.1 m sodium citrate ph 3.5. the crystals diffract x-rays beyond 1.4 a resolution and belong to the orthorhombic space group p2(1)2(1)2(1), with unit-cell parameter ...200010771441
a membrane-bound flavocytochrome c-sulfide dehydrogenase from the purple phototrophic sulfur bacterium ectothiorhodospira vacuolata.the amino acid sequence of ectothiorhodospira vacuolata cytochrome c-552, isolated from membranes with n-butanol, shows that it is a protein of 77 amino acid residues with a molecular mass of 9,041 da. it is closely related to the cytochrome subunit of chlorobium limicola f. sp. thiosulfatophilum flavocytochrome c-sulfide dehydrogenase (fcsd), having 49% identity. these data allowed isolation of a 5.5-kb subgenomic clone which contains the cytochrome gene and an adjacent flavoprotein gene as in ...200010809687
exploitation of butyrate kinase and phosphotransbutyrylase from clostridium acetobutylicum for the in vitro biosynthesis of poly(hydroxyalkanoic acid).active butyrate kinase (buk) and phosphotransbutyrylase (ptb) were purified in three steps: ammonium sulfate precipitation, hydrophobic chromatography on phenyl-sepharose and affinity chromatography on matrex red a from recombinant escherichia coli k2006 (pjc7). they were then successfully exploited for in vitro synthesis of 3-hydroxybutyryl-coa (3hbcoa), 4-hydroxybutyryl-coa (4hbcoa), 4-hydroxyvaleryl-coa (4hvcoa) and poly(hydroxyalkanoic acid) (pha). in addition, the ability of the pha synthas ...200010855714
structural examination of the nickel site in chromatium vinosum hydrogenase: redox state oscillations and structural changes accompanying reductive activation and co binding.an x-ray absorption spectroscopic study of structural changes occurring at the ni site of chromatium vinosum hydrogenase during reductive activation, co binding, and photolysis is presented. structural details of the ni sites for the ready silent intermediate state, si(r), and the carbon monoxide complex, si-co, are presented for the first time in any hydrogenase. analysis of nickel k-edge energy shifts in redox-related samples reveals that reductive activation is accompanied by an oscillation i ...200010858296
complex formation between chromatium vinosum ferric cytochrome c' and bromophenol blue.an unusual complex has been observed between the common electrophoresis tracer bromophenol blue (bpb) and the cytochrome c' from chromatium vinosum during polyacrylamide gel electrophoresis. complex formation results in a shift and increase in the intensity of the visible absorption band of bpb. differential spectrophotometric titration of bpb with cytochrome c' indicates that one bpb binds to each of the two subunits of cytochrome c' with a binding constant of 4.2(0.5) x 10(5). the absence of a ...200010871042
[the activity of the carbon metabolism enzymes in chromatium minutissimum after long-term preservation].the activity of the enzymes of the tricarboxylic acid cycle and glyoxylate shunt, as well as of some enzymes involved in carbohydrate metabolism, were determined in the purple sulfur bacterium chromatium minutissimum, either maintained by subculturing in liquid medium or stored in the lyophilized state for 36 years. in cultures stored in the lyophilized state, the activities of the key enzymes of the tricarboxylic acid cycle, glyoxylate shunt, and embden-meyerhof-parnas pathway were higher, wher ...200010920800
characterization of the cys gene locus from allochromatium vinosum indicates an unusual sulfate assimilation pathway.homologues of the genes cysb, cysi, cysh, cysd, cysn, and seld were identified in the genome of the phototrophic purple sulfur bacterium allochromatium vinosum (formerly chromatium vinosum). on the basis of amino acid comparisons these genes encode a ferredoxin-dependent siroheme-sulfite reductase (cysi), a plant-type assimilatory aps reductase without thioredoxin domain (cysh), the two different subunits of heterodimeric atp sulfurylase (cysdn), a transcriptional regulator (cysb) and a selenoph ...200010939523
model-free analysis of a thermophilic fe(7)s(8) protein compared with a mesophilic fe(4)s(4) protein.15n t(1), t(2) and (1)h-(15)n noe were measured for the thermophilic fe(7)s(8) protein from bacillus schlegelii and for the fe(4)s(4) hipip protein from chromatium vinosum, which is a mesophilic protein. the investigation was performed at 276, 300, and 330 k at 11.7 t for the former, whereas only the 298 k data at 14.1 t for the latter were acquired. the data were analyzed with the model-free protocol after correcting the measured parameters for the effect of paramagnetism, because both proteins ...200010944395
analysis of the thiocapsa pfennigii polyhydroxyalkanoate synthase: subcloning, molecular characterization and generation of hybrid synthases with the corresponding chromatium vinosum enzyme.the pha synthase structural gene of thiocapsa pfennigii was identified and subcloned on a 2.8-kbp bamhi restriction fragment, which was cloned recently from a genomic 15.6-kbp ecori restriction fragment. nucleotide sequence analysis of this fragment revealed three open reading frames (orfs), representing coding regions. two orfs encoded for the phae (mr 40,950) and phac (mr 40,190) subunits of the pha synthase from t. pfennigii and exhibited high homology with the corresponding proteins of the c ...200010968631
the role of structural intersubunit microheterogeneity in the regulation of the activity in hysteresis of ribulose 1, 5-bisphosphate carboxylase/oxygenase.many enzymes are composed of subunits with the identical primary structure. it has been believed that the protein structure of these subunits is the same. ribulose 1,5-bisphosphate carboxylase/oxygenase (rubisco) comprises eight large subunits with the identical amino acid sequence and eight small subunits. rotation of the side chains of the lysine residues, lys-21 and lys-305, in each of the eight large subunits in spinach rubisco in two ways produces microheterogeneity among the subunits. thes ...200011011141
energy transfer and charge separation in the purple non-sulfur bacterium roseospirillum parvum.the antenna reaction centre system of the recently described purple non-sulfur bacterium roseospirillum parvum strain 930i was studied with various spectroscopic techniques. the bacterium contains bacteriochlorophyll (bchl) a, 20% of which was esterified with tetrahydrogeranylgeraniol. in the near-infrared, the antenna showed absorption bands at 805 and 909 nm (929 nm at 6 k). fluorescence bands were located at 925 and 954 nm, at 300 and 6 k, respectively. fluorescence excitation spectra and tim ...200011106774
mechanistic studies on class i polyhydroxybutyrate (phb) synthase from ralstonia eutropha: class i and iii synthases share a similar catalytic mechanism.the class i and iii polyhydroxybutyrate (phb) synthases from ralstonia eutropha and chromatium vinosum, respectively, catalyze the polymerization of beta-hydroxybutyryl-coenzyme a (hbcoa) to generate phb. these synthases have different molecular weights, subunit composition, and kinetic properties. recent studies with the c. vinosum synthase suggested that it is structurally homologous to bacterial lipases and allowed identification of active site residues important for catalysis [jia, y., kappo ...200111170423
model of detergent-induced spectral changes of the b800-850 complex from chromatium minutissimum.the absorption and circular dichroism spectra of the b800-850 complex from chromatium minutissimum before and after the triton x-100 treatment were simulated by means of standard exciton theory, taking into account inhomogeneous broadening. to explain the spectral changes of the b800-850 complex treated with triton x-100, we have assumed that all bacteriochlorophyll pigments absorbing at 850 nm exhibit the same additional rotation of approximately 20 degrees around the axis perpendicular to the ...200011185955
comparison and characterization of the [fe4s4]2+/3+ centre in the wild-type and c77s mutated hipips from chromatium vinosum monitored by mössbauer, 57fe endor and epr spectroscopies.mössbauer, 57fe endor, cw and pulsed epr experiments were performed on the reduced and the oxidized high-potential iron proteins (hipips) of the wild type (wt) and the c77s mutant from chromatium vinosum. the epr spectra of the oxidized wt and mutant show three species respectively having nearly the same g-values but strongly changed spectral contributions. relaxation times were estimated for oxidized wt and mutant at t = 5 k with pulsed epr. a-tensor components of both iron pairs were obtained ...200111315559
elucidation of a [4fe-4s] cluster degradation pathway: rapid kinetic studies of the degradation of chromatium vinosum hipip.irreversible disassembly of the 4fe-4s cluster in chromatium vinosum high-potential iron protein (hipip) has been investigated in the presence of a low concentration of guanidinium hydrochloride. from the dependence of degradation rate on [h+], it is deduced that at least three protons are required to trigger efficient cluster degradation. under these conditions the protonated cluster shows broadened mössbauer signals, but delta eq (1.1 mm/s) and delta (0.44 mm/s) are similar to the native form. ...200111315562
intramolecular electron transfer in [4fe-4s] proteins: estimates of the reorganization energy and electronic coupling in chromatium vinosum ferredoxin.the semi-classical electron transfer theory has been very successful in describing reactions occurring in biological systems, but the relevant parameters in the case of iron-sulfur proteins remain unknown. the recent discovery that 2[4fe-4s] proteins homologous to chromatium vinosum ferredoxin contain clusters with different reduction potentials now gives the opportunity to study the dependence of the intramolecular electron transfer rate between these clusters as a function of the driving force ...200111372203
characterization of clutathione amide reductase from chromatium gracile. identification of a novel thiol peroxidase (prx/grx) fueled by glutathione amide redox cycling.among the chromatiaceae, the glutathione derivative gamma-l-glutamyl-l-cysteinylglycine amide, or glutathione amide, was reported to be present in facultative aerobic as well as in strictly anaerobic species. the gene (garb) encoding the central enzyme in glutathione amide cycling, glutathione amide reductase (gar), has been isolated from chromatium gracile, and its genomic organization has been examined. the garb gene is immediately preceded by an open reading frame encoding a novel 27.5-kda ch ...200111399772
first observation by mass spectrometry of a 3+ oxidation state for a [4fe-4s] metalloprotein: an esi-fticr mass spectrometry study of the high potential iron-sulfur protein from chromatium vinosum.electrospray ionization (esi) fourier transform ion cyclotron resonance mass spectrometry (fticr) is used to measure the molecular weight of the high potential iron-sulfur protein (hipip) from chromatium vinosum (c. vinosum) and its corresponding apoprotein. by accurate mass measurement of the metalloprotein, the oxidation state of the [4fe-4s] metal center is assigned as 3+. this is the highest oxidation state yet observed by mass spectrometry for a [4fe-4s] cluster, which usually appears in th ...200111444604
uv-induced destruction of light-harvesting complexes from purple bacterium chromatium minutissimum.we studied uv-induced photodestruction of the native forms of bacteriochlorophyll a (bchl a) from chromatophores and light harvesting complexes (lhc) of the sulphur photosynthetic bacterium chromatium minutissimum. irradiation of chromato- phores with 365-nm light (soret band) or 280-nm light (absorption region of aromatic amino acids) led to the destruction of all long-wavelength forms of bchl a. the quantum yields of photodestruction produced by the 280-nm light was higher than that produced b ...200111497101
the stratified microbial community at laguna figueroa, baja california, mexico: a possible model for prephanerozoic laminated microbial communities preserved in cherts.the microbial mat community of the evaporite flat at north pond, laguna figueroa (baja california, mexico) was actively involved in the production of laminated sediments prior to 1978. heavy rains in 1979 and 1980 flooded the mat with 1 and 3 meters of meteoric water respectively. the flooding deposited up to 10 cm of silicoclastic sediment over the microcoleus-dominated mat and resulted in the cessation of laminated sediment deposition. in 1982, the surface had been recolonized by species of ...198411536585
contrasting modes of photosynthetic enzyme regulation in oxygenic and anoxygenic prokaryotes.enzymes that are regulated by the ferredoxin/thioredoxin system in chloroplasts--fructose-1,6-bis-phosphatase (fbpase), sedoheptulose-1,7-bisphosphatase (sbpase), and phosphoribulokinase (prk)--were partially purified from two different types of photosynthetic prokaryotes (cyanobacteria, purple sulfur bacteria) and tested for a response to thioredoxins. each of the enzymes from the cyanobacterium nostoc muscorum, an oxygenic organism known to contain the ferredoxin/thioredoxin system, was activ ...198411536590
carbon isotope fractionation by thermophilic phototrophic sulfur bacteria: evidence for autotrophic growth in natural populations.purple phototrophic bacteria of the genus chromatium can grow as either photoautotrophs or photoheterotrophs. to determine the growth mode of the thermophilic chromatium species, chromatium tepidum, under in situ conditions, we have examined the carbon isotope fractionation patterns in laboratory cultures of this organism and in mats of c. tepidum which develop in sulfide thermal springs in yellowstone national park. isotopic analysis (13c/12c) of total carbon, carotenoid pigments, and bacteri ...198911536609
the microbial community at laguna figueroa, baja california mexico: from miles to microns.laguna figueroa is a lagoonal complex on the pacific coast of the baja california penisula 200 km south of the mexican-united states border. the hypersaline lagoon is 16 km long and 2-3 km wide with a salt marsh and evaporite flat and is separated from the ocean by a barrier dune and beach. at the salt marsh-evaporite flat interface a stratified microbial community dominated by microcoleus chthonoplastes is depositing laminated sediments. similar stratiform deposits with associated microbial ...198511539612
isotope effects associated with the anaerobic oxidation of sulfite and thiosulfate by the photosynthetic bacterium, chromatium vinosum.the purple photosynthetic bacterium chromatium vinosum, strain d, catalyzes several oxidations of reduced sulfur compounds under anaerobic conditions in the light: e.g., sulfide --> sulfur --> sulfate, sulfite --> sulfate, and thiosulfate --> sulfur + sulfate. here it is shown that no sulfur isotope effect is associated with the last of these processes; isotopic compositions of the sulfur and sulfate produced can differ, however, if the sulfane and sulfonate positions within the thiosulfate ha ...198511540842
predatory prokaryotes: predation and primary consumption evolved in bacteria.two kinds of predatory bacteria have been observed and characterized by light and electron microscopy in samples from freshwater sulfurous lakes in northeastern spain. the first bacterium, named vampirococcus, is gram-negative and ovoidal (0.6 micrometer wide). an anaerobic epibiont, it adheres to the surface of phototrophic bacteria (chromatium spp.) by specific attachment structures and, as it grows and divides by fission, destroys its prey. an important in situ predatory role can be inferr ...198611542073
the evolution of glutathione metabolism in phototrophic microorganisms.of the many roles ascribed to glutathione (gsh) the one most clearly established is its role in the protection of higher eucaryotes against oxygen toxicity through destruction of thiol-reactive oxygen byproducts. if this is the primary function of gsh then gsh metabolism should have evolved during or after the evolution of oxygenic photosynthesis. that many bacteria do not produce gsh is consistent with this view. in the present study we have examined the low-molecular-weight thiol compositio ...198711542078
competition for sulfide among colorless and purple sulfur bacteria in cyanobacterial mats.the vertical zonation of light, o2, h2s, ph, and sulfur bacteria was studied in two benthic cyanobacterial mats from hypersaline ponds at guerrero negro, baja california, mexico. the physical-chemical gradients were analyzed in the upper few mm at < or = 100 micrometers spatial resolution by microelectrodes and by a fiber optic microprobe. in mats, where oxygen produced by photosynthesis diffused far below the depth of the photic zone, colorless sulfur bacteria (beggiatoa sp.) were the dominan ...198611542103
steady-state and time-resolved fluorescence studies on wild type and mutant chromatium vinosum high potential iron proteins: holo- and apo-forms.detailed circular dichroism (cd), steady-state and time-resolved tryptophan fluorescence studies on the holo- and apo- forms of high potential iron protein (hipip) from chromatium vinosum and its mutant protein have been carried out to investigate conformational properties of the protein. cd studies showed that the protein does not have any significant secondary structure elements in the holo- or apo- hipip, indicating that the metal cluster does not have any effect on formation of secondary str ...200111566801
spectroscopic and functional properties of novel 2[4fe-4s] cluster-containing ferredoxins from the green sulfur bacterium chlorobium tepidum.two distinct ferredoxins, fd i and fd ii, were isolated and purified to homogeneity from photoautotrophically grown chlorobium tepidum, a moderately thermophilic green sulfur bacterium that assimilates carbon dioxide by the reductive tricarboxylic acid cycle. both ferredoxins serve a crucial role as electron donors for reductive carboxylation, catalyzed by a key enzyme of this pathway, pyruvate synthase/pyruvate ferredoxin oxidoreductase. the reduction potentials of fd i and fd ii were determine ...200111568186
evaluation of solvent accessibility to the [fe(4)s(4)] binding pocket in native and tyr19 mutant high potential iron proteins by (1)h-(15)n hmqc and (19)f nmr experiments.the solvent accessibility of chromatium vinosumhigh potential iron protein (hipip) has been investigated by use of (1)h-(15)n hmqc, and (19)f nmr spectroscopy. these nmr experiments indicate that solvent accessibility to the cluster core is similar, and minimal, for the reduced and oxidized states of native hipip, but increases significantly for mutant proteins (tyr19leu and tyr19his). these results support a proposed role [agarwal, a.; li, d.; cowan, j. a. proc. natl. acad. sci. u.s.a. 1995, 92 ...199611666298
azotobacter vinelandii ferredoxin i: a sequence and structure comparison approach to alteration of [4fe-4s]2+/+ reduction potential.the reduction potential (e(0)') of the [4fe-4s](2+/+) cluster of azotobacter vinelandii ferredoxin i (avfdi) and related ferredoxins is approximately 200 mv more negative than the corresponding clusters of peptostreptococcus asaccharolyticus ferredoxin and related ferredoxins. previous studies have shown that these differences in e(0)' do not result from the presence or absence of negatively charged surface residues or in differences in the types of hydrophobic residues found close to the [4fe-4 ...200211704670
the [nife] hydrogenase from allochromatium vinosum studied in epr-detectable states: h/d exchange experiments that yield new information about the structure of the active site.in this study we report on thus-far unobserved proton hyperfine couplings in the well-known epr signals of [nife] hydrogenases. the preparation of the enzyme in several highly homogeneous states allowed us to carefully re-examine the ni(u)*, ni(r)*, ni(a)-c* and ni(a)-l* epr signals which are present in most [nife] hydrogenases. at high resolution (modulation amplitude 0.57 g), clear indications for hyperfine interactions were observed in the g(z) line of the ni(r)* epr signal. the hyperfine pat ...200111713683
crystal structure and stability studies of c77s hipip: a serine ligated [4fe-4s] cluster.the crystal structure of chromatium vinosum c77s hipip has been determined and is compared with that of wild type. this is the first reported crystal structure of a ser ligated [4fe-4s] cluster and reveals a 0.11 a shortening of the fe-o bond (relative to fe-s), but only minor structural alterations of the overall tertiary structure. coordination changes are corroborated by resonance raman spectroscopy. comparison of the crystal and solution structures for hipips identifies phe48 as the main con ...200211802718
crystallization and preliminary x-ray crystallographic analysis of glutathione amide reductase from chromatium gracile.the chromatiaceae-specific glutathione amide reductase (gar) belongs to the well known family of the glutathione reductases, even though differences in both substrate (glutathione amide instead of glutathione) and coenzyme (nadh instead of nadph) specificities are reported. crystals of the gar enzyme from chromatium gracile have been grown at 294 k by the hanging-drop vapour-diffusion method using lithium sulfate as a precipitant in the presence of nickel ions. the crystals belong to space group ...200211807270
effect of carotenoids on the interaction between pigment-protein complexes in membranes of the sulfur photosynthetic bacterium chromatium minutissimum. 200111813560
redox-related chemical shift perturbations on backbone nuclei of high-potential iron-sulfur proteins. 200211896740
a coupling factor from chromatium strain d chromatophores. 197111945626
n-terminal methylation of the core light-harvesting complex in purple photosynthetic bacteria.several core light-harvesting complexes from both sulfur and non-sulfur purple photosynthetic bacteria have been identified to be methylated at the n-terminal alpha-amino group of beta-polypeptides by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nuclear magnetic resonance. monomethylation has been confirmed for the n-terminal alanine residues of the beta-polypeptides from rhodospirillum rubrum, thermochromatium tepidum and chromatium vinosum, but not for ...200212023037
the role of high-potential iron protein and cytochrome c(8) as alternative electron donors to the reaction center of chromatium vinosum.under anaerobic conditions, intact cells of the purple sulfur bacterium chromatium vinosum exhibit rapid photooxidation of the two low-potential hemes of the c-type cytochrome associated with the reaction center, after exposure to two short light flashes separated by a dark interval. reduction of the photooxidized low-potential hemes is very slow under these conditions. on subsequent flashes, rapid photooxidation of a high-potential reaction center heme occurs and is followed by its rereduction ...200212102629
genetic construction of truncated and chimeric metalloproteins derived from the alpha subunit of acetyl-coa synthase from clostridium thermoaceticum.in this study, a genetics-based method is used to truncate acetyl-coenzyme a synthase from clostridium thermoaceticum (acs), an alpha(2)beta(2) tetrameric 310 kda bifunctional enzyme. acs catalyzes the reversible reduction of co(2) to co and the synthesis of acetyl-coa from co (or co(2) in the presence of low-potential reductants), coa, and a methyl group bound to a corrinoid-iron sulfur protein (cofesp). acs contains seven metal-sulfur clusters of four different types called a, b, c, and d. the ...200212121109
direct comparison of the electrocatalytic oxidation of hydrogen by an enzyme and a platinum catalyst.it is shown that for molecules of allochromatium vinosum [nife]-hydrogenase adsorbed on a pyrolytic graphite electrode the nickel-iron active site catalyzes hydrogen oxidation at a diffusion-controlled rate matching that achieved by platinum.200212123018
dynamics of wild-type hipips: a cys77ser mutant and a partially unfolded hipip.the temperature dependence of the mean square displacement of the (57)fe nuclei due to motion faster than 100 ns are measured by temperature-dependent mössbauer spectroscopy for oxidized and reduced hipips from ectothiorhodospira halophila, chromatium vinosum wt and a cys77ser mutant. the behaviour is interpretable in the frame of the general model of protein dynamics distinguishing two temperature intervals. the character of harmonic and quasi-diffusional modes in hipips is discussed. dynamic i ...200212203006
cloning a gene encoding a light-harvesting i polypeptide from ectothiorhodospira sp.trying to detect the genes coding for light harvesting ii polypeptides of the purple bacteria ectothiorhodospira sp. a gene corresponding to a light harvesting i polypeptide was cloned. this paper discusses the probable reasons of this result. the sequence of this polypeptide underlines the strong similarity with light harvesting complexes from bacteria such as rhodospirillum molischianum and chromatium vinosum.200212240998
mechanism and biological role of nitric oxide binding to cytochrome c'.the binding of nitric oxide to ferric and ferrous chromatium vinosum cytochrome c' was studied. the extinction coefficients for the ferric and ferrous nitric oxide complexes were measured. a binding model that included both a conformational change and dissociation of the dimer into subunits provided the best fit for the ferric cytochrome c' data. the no (nitric oxide) binding affinity of the wt ferric form was found to be comparable to the affinities displayed by the ferric myoglobins and hemogl ...200212269801
fluorescence of native and carotenoid-depleted lh2 from chromatium minutissimum, originating from simultaneous two-photon absorption in the spectral range of the presumed (optically 'dark') s(1) state of carotenoids.native and carotenoid-depleted peripheral purple bacterial light-harvesting complex (lh2) were investigated by simultaneous two-photon excited (between 1300-1500 nm) fluorescence (tpf). tpf results from direct bacteriochlorophyll excitation in both samples. the spectral position of the 2a(g)(-) state of rhodopin [corrected] is indicated by a diminuition of the bacteriochlorophyll tpf in native lh2. in conclusion, comparison to carotenoid-depleted samples is a conditio sine qua non for unambiguou ...200212297310
predatory prokaryotes: an emerging research opportunity.predatory prokaryotes have evolved a unique strategy of obtaining energy and biosynthetic materials from their surroundings: acquiring them from other living bacterial cells. these types of microbes have been found in a diverse variety of environments, and may play an important role in modulating microbial population structure and dynamics, as has been hypothesized for marine viruses and possibly protists. only one genus of predatory bacterium, bdellovibrio, has been extensively described and st ...200212432957
[two-photon excitation fluorescence spectrum of the light-harvesting complex lh2 from chromatium minutissimum within 650-745 nm range is determined by two-photon absorption of bacteriochlorophyll rather than of carotenoids].two-photon fluorescence excitation spectra of the peripheral light-harvesting complex lh2 from the purple photosynthetic bacterium chromatium minutissimum were examined within the expected spectral range of the optically forbidden s1 singlet state of carotenoids. lh2 preparations isolated from wild-type and carotenoid-depleted cells were used. 100-fs laser pulses in the range of 1300-1490 nm with an energy of 7-9 mw (corresponding to one-photon absorption between 650 and 745 nm) were used for tw ...200212500564
the crystal structure of a 26-nucleotide rna containing a hook-turn.a crystal structure has been obtained for a 26-nucleotide rna that contains the loop e sequence from chromatium minutissimum. rather than having a loop e-like conformation, it consists of an a-form helix that splits into two separate strands following a sheared a-g base pair. the backbone of the strand containing the g of the a-g pair makes a turn of almost 180 degrees in the space of two nucleotides, and then interacts with the minor groove of the helix from which it originates. similar structu ...200312554875
[light-dependent incorporation of selenite into selenocysteine by isolated chromatophore of chromatium vinosum].illiminated intact chromatophore of chromatium vinosum in the presence of o-acetylserine(oas) catalysed incorporation of seo3(2-) into selenocysteine at rate of 359 nmol.mgbchl-1.h-1. sonicated chromatophore catalysed seo3(2-) incorporation at 1.1% of the rate of intact chromatophore. addition of gsh and nadph increased the rate to 88.3% of intact rate, but seo3(2-) incorporation under these conditions was essentially light dependent. the purified gsh reductase from chromatium vinosum in the pre ...199912555543
reconstitution of carotenoids into the light-harvesting complex b800-850 of chromatium minutissimum.chromatophores and peripheral light-harvesting complexes b800-850 with a trace of carotenoids were isolated from chromatium minutissimum cells in which carotenoid biosynthesis was inhibited by diphenylamine. three methods previously used for the reconstitution of carotenoids into either the light-harvesting (lh1) type complexes or reaction centers (rc) of carotenoidless mutants were examined for the possibility of carotenoid reconstitution into the carotenoid depleted chromatophores. all these m ...200312948391
nitrogen fixation and photoproduction of molecular hydrogen by thiorhodaceae. 195313041177
hematin compounds in photosynthetic bacteria. 195413221572
enzymatic activities affecting cytochromes in photosynthetic bacteria. 195413221573
chromatium cytochrome. 195513259698
light-induced uptake of inorganic phosphate in cell-free extracts of obligately anaerobic photosynthetic bacteria. 195613315332
fluorescence of suspensions of green sulphur bacteria. 195613315333
the carotenoids of photosynthetic bacteria. i. the nature of the carotenoid pigments in a halophilic photosynthetic sulphur bacterium (chromatium spp.). 195613340831
chromatium cytochrome. 195613363861
ph-induced changes of the infrared absorption spectra of purple bacteria. 195613373840
observations on the infrared absorption spectrum of bacteriochlorophyll. 195613373841
[studies on assimilation of free carbonic acid by purple sulfur bacteria in the lake belovod]. 195713440598
composition of the photoactive subcellular particles from chromatium. 195713459446
photosynthetic activity of isolated chloroplast fragments of spirogyra. 195713479414
photophosphorylation by subcellular particles from chromatium. 195713479415
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