Methanol:coenzyme M methyltransferase from Methanosarcina barkeri -- substitution of the corrinoid harbouring subunit MtaC by free cob(I)alamin.
Methyl-coenzyme M formation from coenzyme M and methanol in Methanosarcina barkeri is catalysed by an enzyme system composed of three polypeptides MtaA, MtaB and MtaC, the latter of which harbours a corrinoid prosthetic group. We report here that MtaC can be substituted by free cob(I)alamin which is methylated with methanol in an MtaB-catalysed reaction and demethylated with coenzyme M in an MtaA-catalysed reaction. Methyl transfer from methanol to coenzyme M was found to proceed at a relatively high specific activity at micromolar concentrations of cob(I)alamin. This finding was surprising because the methylation of cob(I)alamin catalysed by MtaB alone and the demethylation of methylcob(III)alamin catalysed by MtaA alone exhibit apparent Km for cob(I)alamin and methylcob(III)alamin of above 1 mm. A possible explanation is that MtaA positively affects the MtaB catalytic efficiency and vice versa by decreasing the apparent Km for their corrinoid substrates. Activation of MtaA by MtaB was methanol-dependent. In the assay for methanol:coenzyme M methyltransferase activity cob(I)alamin could be substituted by cob(I)inamide which is devoid of the nucleotide loop. Substitution was, however, only possible when the assays were supplemented with imidazole: approximately 1 mm imidazole being required for half-maximal activity. Methylation of cob(I)inamide with methanol was found to be dependent on imidazole but not on the demethylation of methylcob(III)inamide with coenzyme M. The demethylation reaction was even inhibited by imidazole. The structure and catalytic mechanism of the MtaABC complex are compared with the cobalamin-dependent methionine synthase.
Sauer
K
K
Max-Planck-Institut für terrestrische Mikrobiologie, Philipps- Universität, Marburg, Germany.
Thauer
R K
RK
D002384:Catalysis; D003038:Cobamides / Q000737:chemistry; D045728:Corrinoids; D007093:Imidazoles / Q000737:chemistry; D017021:Methanosarcina barkeri / Q000201:enzymology*; D008780:Methyltransferases / Q000737:chemistry / Q000378:metabolism*; D011166:Porphyrins / Q000737:chemistry*; D014805:Vitamin B 12 / Q000737:chemistry*
D003038:Cobamides; D045728:Corrinoids; D007093:Imidazoles; D011166:Porphyrins; C017142:cobinamide; D008780:Methyltransferases; C036890:methanol-2-mercaptoethanesulfonic acid methyltransferase; D014805:Vitamin B 12