Characterization of a molybdenum cofactor biosynthetic gene cluster in Rhodobacter capsulatus which is specific for the biogenesis of dimethylsulfoxide reductase.
The DMSO reductase of Rhodobacter capsulatus contains a pterin molybdenum cofactor (Moco) and is located in the periplasm. DNA sequence analysis identified four genes involved in the biosynthesis of the Moco (moaA, moaD, moeB and moaC) immediately downstream of the dor (DMSO respiratory) gene cluster. Rhodobacter capsulatus MoaA was expressed in Escherichia coli as a His6-tagged protein. Although, the expressed protein formed inclusion bodies, EPR spectroscopy showed that MoaA contains a [3Fe-4S] cluster. A moaA mutant was constructed and its phenotype indicates that the Moco biosynthetic gene cluster downstream of the dor operon is specific for the biogenesis of DMSO reductase. Two forms of DMSO reductase were purified by immunoaffinity chromatography from the moaA mutant. A mature form of DMSO reductase was located in the periplasm and a precursor form was found in the cytoplasm.
Solomon
Peter S
PS
Department of Journal of General Microbiology, The University of Queensland, Brisbane 4072, Australia.
Shaw
Anthony L
AL
Department of Journal of General Microbiology, The University of Queensland, Brisbane 4072, Australia.
Lane
Ian
I
Centre for Magnetic Resonance, The University of Queensland, Brisbane 4072, Australia.
Hanson
Graeme R
GR
Centre for Magnetic Resonance, The University of Queensland, Brisbane 4072, Australia.
Palmer
Tracy
T
Nitrogen Fixation Laboratory, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK.
McEwan
Alastair G
AG
Department of Journal of General Microbiology, The University of Queensland, Brisbane 4072, Australia.
D000595:Amino Acid Sequence; D002874:Chromosome Mapping; D003067:Coenzymes; D004926:Escherichia coli; D005798:Genes, Bacterial; D007506:Iron-Sulfur Proteins; D008667:Metalloproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D008982:Molybdenum / Q000378:metabolism; D000090006:Molybdenum Cofactors; D005810:Multigene Family; D009154:Mutation; D010088:Oxidoreductases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D011621:Pteridines / Q000378:metabolism*; D016354:Rhodobacter capsulatus / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D003067:Coenzymes; D007506:Iron-Sulfur Proteins; D008667:Metalloproteins; D008982:Molybdenum; D000090006:Molybdenum Cofactors; D010088:Oxidoreductases; D011621:Pteridines; C022261:molybdopterin; C045664:dimethyl sulfoxide reductase