Abstract
Dissimilatory sulfite reductase (DsrAB) of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough is an α2β2 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.
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References
Aketagawa J, Kojo K & Ishimoto M (1985) Purification and properties of sulfite reductase from Desulfovibrio vulgaris Miyazaki F. Agric. Biol. Chem. 49: 2359–2365
Dahl C, Kredich NM, Deutzmann R & Trüper HG (1993) Dissimilatory sulphite reductase from Archaeoglobus fulgidus: physicochemical properties of the enzyme and cloning, sequencing and analysis of the reductase genes. J. Gen. Microbiol. 139: 1817–1828
Deléage G, Blanchet C, & Geourjon C (1997) Protein structure prediction. Implications for the biologist. Biochimie 79: 681–686
Hipp WM, Pott AS, Thum-Schmitz N, Faath I, Dahl C & Trüper HG (1997) Towards the phylogeny of APS reductases and sirohaem sulfite reductases in sulfate-reducing and sulfite-oxidizing prokaryotes. Microbiology 143: 2891–2902
Karkhoff-Schweizer RR, Bruschi M & Voordouw G (1993) Expression of the γ-subunit gene of desulfoviridin-type dissimilatory sulfite reductase and of the α-and β-subunit genes is not coordinately regulated. Eur. J. Biochem. 211: 501–507
Karkhoff-Schweizer RR, Huber DPW & Voordouw G (1995) Conservation of the genes for dissimilatory sulfite reductase from Desulfovibrio vulgaris and Archaeoglobus fulgidus allows their detection by PCR. Appl. Environ. Microbiol. 61: 290–296
Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680–685
Larsen O, Lien T & Birkeland NK (1999) Dissimilatory sulfite reductase from Archaeoglobus profundus and Desulfotomaculum thermocisternum: phylogenetic and structural implications from gene sequences. Extremophiles 3: 63–70
Matsudaira P (1987) Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J. Biol. Chem. 262: 10035–10038
Molitor M, Dahl C, Molitor I, Schafer U, Speich N, Huber R, Deutzmann R & Trüper HG (1998) A dissimilatory sirohaemsulfite-reductase-type protein from the hyperthermophilic archaeon Pyrobaculum islandicum. Microbiology 144: 529–541
Peck Jr HD (1993) Bioenergetic strategies of the sulfate-reducing bacteria. In: Odom JM & Singleton Jr R (Ed.) The sulfatereducing bacteria: contemporary perspectives (pp 41–76). Springer-Verlag, New York
Pierik AJ, Duyvis MG, van Helvoort JMLM, Wolbert RBG & Hagen WR (1992) The third subunit of desulfoviridin-type dissimilatory sulfite reductase. Eur. J. Biochem. 205: 111–115
Pollock WBR, Chemerika PJ, Forrest ME, Beatty JT & Voordouw G (1989) Expression of the gene encoding cytochrome c3 from Desulfovibrio vulgaris (Hildenborough) in Escherichia coli: export and processing of the apoprotein. J. Gen. Microbiol. 135: 2319–2328
Postgate JR (1984) The sulphate-reducing bacteria, 2nd edn. Cambridge University Press, Cambridge
Sambrook J, Fritsch EF & Maniatis T (1989) Molecular cloning: a laboratory manual. 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Singleton, R. Jr. (1993) The sulfate-reducing bacteria: an overview. In: Odom JM & Singleton Jr R (Eds) The sulfate-reducing bacteria: contemporary perspectives (pp 1–19). Springer-Verlag, New York
Steuber J, Arendsen AF, Hagen WR & Kroneck PMH (1995) Molecular properties of the dissimilatory sulfite reductase from Desulfovibrio desulfuricans (Essex) and comparison with the enzyme from Desulfovibrio vulgaris (Hildenborough). Eur. J. Biochem. 233: 873–879
Tabor S (1991) Expression using the T7 RNA polymerase/promoter system. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Smith JA & Struhl K (Eds). Current protocols in molecular biology (pp 16.2.1–16.2.11). Green Publishing Associates and Wiley & Sons Inc., New York
Towbin H, Staehelin T & Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76: 4350–4354
Voordouw G, Strang JD & Wilson FR (1989) Organization of the genes encoding [Fe] hydrogenase in Desulfovibrio vulgaris subsp. oxamicus Monticello. J. Bacteriol. 171: 3881–3889
Wagner M, Roger AJ, Flax JL, Brusseau GA & Stahl DA (1998) Phylogeny of dissimilatory sulfite reductases supports an early origin of sulfate respiration. J. Bacteriol. 180: 2975–2982
Yanisch-Perron C, Vieira J & Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33: 103–119
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Hittel, D.S., Voordouw, G. Overexpression, purification and immunodetection of DsrD from Desulfovibrio vulgaris Hildenborough. Antonie Van Leeuwenhoek 77, 271–280 (2000). https://doi.org/10.1023/A:1002449227469
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DOI: https://doi.org/10.1023/A:1002449227469