Evidence for the physiological role of a rhodanese-like protein for the biosynthesis of the molybdenum cofactor in humans.
Recent studies have identified the human genes involved in the biosynthesis of the molybdenum cofactor. The human MOCS3 protein contains an N-terminal domain similar to the Escherichia coli MoeB protein and a C-terminal segment displaying similarities to the sulfurtransferase rhodanese. The MOCS3 protein is believed to catalyze both the adenylation and the subsequent generation of a thiocarboxylate group at the C terminus of the smaller subunit of molybdopterin (MPT) synthase. The MOCS3 rhodanese-like domain (MOCS3-RLD) was purified after heterologous expression in E. coli and was shown to catalyze the transfer of sulfur from thiosulfate to cyanide. In a defined in vitro system for the generation of MPT from precursor Z, the sulfurated form of MOCS3-RLD was able to provide the sulfur for the thiocarboxylation of MOCS2A, the small MPT synthase subunit in humans. Mutation of the putative persulfide-forming active-site cysteine residue C412 abolished the sulfurtransferase activity of MOCS3-RLD completely, showing the importance of this cysteine residue for catalysis. In contrast to other mammalian rhodaneses, which are mostly localized within mitochondria, MOCS3 in addition to the subunits of MPT synthase are localized in the cytosol.
Matthies
Andreas
A
Department of Plant Biology, Technical University Braunschweig, 38023 Braunschweig, Germany.
Rajagopalan
K V
KV
Mendel
Ralf R
RR
Leimkühler
Silke
S
D001483:Base Sequence; D003067:Coenzymes / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D008667:Metalloproteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D000090006:Molybdenum Cofactors; D016297:Mutagenesis, Site-Directed; D011621:Pteridines / Q000378:metabolism; D013884:Thiosulfate Sulfurtransferase / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
D003067:Coenzymes; D017931:DNA Primers; D008667:Metalloproteins; D000090006:Molybdenum Cofactors; D011621:Pteridines; D013884:Thiosulfate Sulfurtransferase; C022261:molybdopterin