Cloning and sequencing of the gene for rubrerythrin from Desulfovibrio vulgaris (Hildenborough)

Prickril, Benet C.; Kurtz, Donald M.; LeGall, Jean; Voordouw, Gerrit

doi:10.1021/bi00110a014

Cited by 38 publications

(35 citation statements)

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“…The group 111 residues are organized as follows : -C-X,-C-X,,-,,-C-P-X-C-(117-160 in H y f H and 117-158 in HycF), and could therefore serve as ligands for a third iron site. A similar motif is found in rubredoxins and rubrerythrins where the cysteine residues ligate a mononuclear iron atom (-C-X,-C-XI,-,,-C-P-X-C-; Prickril et al, 1991 members of the complex I 2 3 kDa-subunit family and a mononuclear site (or iron-sulphur cluster) not present in other family members. However, subsequent studies revealed that the phenotype ascribed to the hycH mutant was due to a polar effect on the downstream gene encoding HycI, which is now known to be involved in hydrogenase maturation (Rossmann et al, 1995).…”

Section: Features Of the Translation Productsmentioning

confidence: 59%

A 12-cistron Escherichia coli operon (hyf) encoding a putative proton-translocating formate hydrogenlyase system

et al. 1997

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The nucleotide sequence has been determined for a twelve-gene operon of Escherichia coli designated the hyf operon (hyfASCD€FGH/R-focB). The hyf operon is located a t 5508-56.0 min and encodes a putative nine-subunit hydrogenase complex (hydrogenase four or Hyf), a potential formate-and 6%-dependent transcriptional activator, HyfR (related to FhIA), and a possible formate transporter, FocB (related to FocA). Five of the nine Hyf-complex subunits are related to subunits of both the E. coli hydrogenase-3 complex (Hyc) and the proton-translocating NADH : quinone oxidoreductases (complex I and Nuo), whereas two Hyf subunits are related solely to NADH : quinone oxidoreductase subunits. The Hyf components include a predicted 523 residue [Ni-Fe] hydrogenase (large subunit) with an N-terminus (residues homologous to the 30 kDa or NuoC subunit of complex 1. It is proposed that Hyf, in conjunction with formate dehydrogenase H (Fdh-H), forms a hitherto unrecognized respiration-linked proton-translocating formate hydrogenlyase (FHL-2). It is likely that HyfR acts as a formate-dependent regulator of the hyf operon and that FocB provides the Hyf complex with external formate as substrate.

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Section: Features Of the Translation Productsmentioning

confidence: 59%

A 12-cistron Escherichia coli operon (hyf) encoding a putative proton-translocating formate hydrogenlyase system

et al. 1997

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“…The molecular mass of each subunit was estimated to be approximately 21,500 Da (12). The rubrerythrin gene (rbr) of D. vulgaris has been cloned and sequenced (23). The analysis of the C-terminal portion of the amino acid sequence confirmed spectroscopic studies (7) which predicted structural similarities to rubredoxin iron centers, and on this basis, four cysteines probably representing iron ligands could be readily identified (23).…”

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confidence: 94%

“…However, interactions of rubrerythrin with O 2 or its immediate reduction products in vivo cannot be ruled out a priori (10). Tests of rubrerythrin for catalase, superoxide dismutase (SOD), nitrate reductase, phosphatase, and pyrophosphatase activities were reported to be negative (12,21,23).In this paper, we report the isolation and characterization of a rubrerythrin from a species other than D. vulgaris, namely, Clostridium perfringens, and we define a role for its function. Physiological and structural implications of these results are examined.…”

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confidence: 99%

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Rubrerythrin from Clostridium perfringens: cloning of the gene, purification of the protein, and characterization of its superoxide dismutase function

1996

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The food-borne pathogen Clostridium perfringens, which is an obligate anaerobe, showed growth under conditions of oxidative stress. In protein extracts we looked for superoxide dismutase (SOD) activities which might scavenge highly toxic superoxide radicals evolving under such stress conditions. Using the classical assay to detect SOD activity on gels after electrophoresis of C. perfringens proteins, we obtained a pattern of three major bands indicating SOD activity. The protein representing the brightest band was purified by three chromatographic steps. On the basis of 20 amino acids determined from the N terminus of the protein, we designed a degenerate oligonucleotide probe to isolate the corresponding gene. We finally sequenced an open reading frame of 195 amino acids (molecular mass, 21,159 Da) with a strong homology to the Desulfovibrio vulgaris rubrerythrin; therefore, we assumed to have cloned a rubrerythrin gene from C. perfringens, and we named it rbr. The C-terminal region of the newly detected rubrerythrin from C. perfringens contains a characteristic non-heme, non-sulfur iron-binding site -Cys-X-X-Cys-(X) 12 -Cys-X-X-Cys-similar to that found in rubrerythrin from D. vulgaris. In addition, three -Glu-X-X-His-sequences could represent diiron binding domains. We observed SOD activity in extracts of Escherichia coli strains containing the recombinant rbr gene from C. perfringens. A biological function of rubrerythrin as SOD was confirmed with the functional complementation by the rbr gene of an E. coli mutant strain lacking SOD activity. We therefore suppose that rubrerythrin plays a role as a scavenger of oxygen radicals.Rubrerythrin is a non-heme, non-sulfur iron protein that was first isolated from the anaerobic sulfate-reducing bacterium Desulfovibrio vulgaris (Hildenborough) (13). Rubrerythrin was first described as a homodimer containing four iron atoms which are arranged into two rubredoxin-like FeS 4 centers and one hemerythrin-like binuclear iron cluster (13). Later on, six iron atoms per rubrerythrin dimer were reported to be distributed in two mononuclear centers and two binuclear clusters (21). The molecular mass of each subunit was estimated to be approximately 21,500 Da (12). The rubrerythrin gene (rbr) of D. vulgaris has been cloned and sequenced (23). The analysis of the C-terminal portion of the amino acid sequence confirmed spectroscopic studies (7) which predicted structural similarities to rubredoxin iron centers, and on this basis, four cysteines probably representing iron ligands could be readily identified (23). Recently, it has been suggested that the oxidized form of the diiron cluster bears a close structural (but not necessarily functional) resemblance to that of the (-oxo)(-carboxylato) diiron cluster in oxidized ribonucleotide reductase (7). Moreover, the visible absorption, electron paramagnetic resonance, and Mössbauer spectra of the FeS 4 site in rubrerythrin are quite similar to those of rubredoxin in both oxidized and reduced forms of the proteins. The midpoint reductio...

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“…D. vulgaris enzymes that detoxify ROS include superoxide reductase (Sor) (3,13,24,34) and rubrerythrin (Rbr), which has NADH-dependent H 2 O 2 reductase activity (4,22,28). Sor and Rbr reduce superoxide and hydrogen peroxide to water without regeneration of oxygen, a feature that is important for oxygen detoxification in anaerobes (13).…”

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confidence: 99%

Function of Oxygen Resistance Proteins in the Anaerobic, Sulfate-Reducing Bacterium Desulfovibrio vulgaris Hildenborough

Fournier

Zhang²,

Wildschut³

et al. 2003

J Bacteriol

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101

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Two mutant strains of Desulfovibrio vulgaris Hildenborough lacking either the sod gene for periplasmic superoxide dismutase or the rbr gene for rubrerythrin, a cytoplasmic hydrogen peroxide (H 2 O 2 ) reductase, were constructed. Their resistance to oxidative stress was compared to that of the wild-type and of a sor mutant lacking the gene for the cytoplasmic superoxide reductase. The sor mutant was more sensitive to exposure to air or to internally or externally generated superoxide than was the sod mutant, which was in turn more sensitive than the wild-type strain. No obvious oxidative stress phenotype was found for the rbr mutant, indicating that H 2 O 2 resistance may also be conferred by two other rbr genes in the D. vulgaris genome.

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Cloning and sequencing of the gene for rubrerythrin from Desulfovibrio vulgaris (Hildenborough)

Cited by 38 publications

References 32 publications

A 12-cistron Escherichia coli operon (hyf) encoding a putative proton-translocating formate hydrogenlyase system

A 12-cistron Escherichia coli operon (hyf) encoding a putative proton-translocating formate hydrogenlyase system

Rubrerythrin from Clostridium perfringens: cloning of the gene, purification of the protein, and characterization of its superoxide dismutase function

Function of Oxygen Resistance Proteins in the Anaerobic, Sulfate-Reducing Bacterium Desulfovibrio vulgaris Hildenborough

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