Overexpression, purification and immunodetection of DsrD from Desulfovibrio vulgaris Hildenborough.
Dissimilatory sulfite reductase (DsrAB) of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough is an alpha2beta2 tetramer of 180 kDa, encoded by the dsr operon. In addition to the dsrA and dsrB genes, this operon contains a gene (dsrD) encoding a protein of only 78 amino acids. Although, the function of DsrD is currently unknown, the presence of a dsrD gene has been demonstrated in a variety of sulfate-reducing bacteria and archaea. DsrD was expressed in Escherichia coli at a very high level and purified to homogeneity. Protein blotting experiments, using antisera raised against purified DsrD, demonstrated that it is expressed constitutively in D. vulgaris and does not copurify with DsrAB. Spectroscopic analysis of DsrD indicated that it does not bind either sulfite or sulfide, the substrate and product, respectively of the reaction catalyzed by DsrAB. Thus, although the conservation of this protein and its demonstrated presence in D. vulgaris, suggest an essential function in dissimilatory sulfite reduction, this function remains to be elucidated.
Hittel
D S
DS
Department of Biological Sciences, University of Calgary, Calgary, Alberta, T2N 1N4, Canada.
Voordouw
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D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002942:Circular Dichroism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D016969:Desulfovibrio vulgaris / Q000201:enzymology / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D050885:Hydrogensulfite Reductase; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009876:Operon; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013440:Sulfides / Q000737:chemistry / Q000378:metabolism; D013447:Sulfites / Q000737:chemistry / Q000378:metabolism
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D013440:Sulfides; D013447:Sulfites; D050862:Oxidoreductases Acting on Sulfur Group Donors; D050885:Hydrogensulfite Reductase