Incorporation of coenzyme M into component C of methylcoenzyme M methylreductase during in vitro methanogenesis

• Published in: The Journal of biological chemistry 1987-04, Vol.262 (12), p.5581-5586
• Main Authors: Hartzell, P.L., Donnelly, M.I., Wolfe, R.S.
• Format: Article
• Language: English
• Subjects: Bacteriology; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; coenzyme M; Euryarchaeota - enzymology; Fundamental and applied biological sciences. Psychology; Kinetics; Mercaptoethanol - analogs & derivatives; Mesna - isolation & purification; Mesna - metabolism; Metabolism. Enzymes; Methane - metabolism; methanogenesis; methyl-coenzyme M reductase; Microbiology; Mission oriented research; Oxidoreductases - isolation & purification; Oxidoreductases - metabolism; Physiology and metabolism; Sulfur Radioisotopes
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)45612-8

Reduction of the methyl group of [methyl-3H,thio-35S]2-methylthioethanesulfonic acid to methane by a reconstituted enzyme system resulted in a slow incorporation of [thio-35S]2-mercaptoethanesulfonic acid (HS-CoM) into component C of the methylreductase system. Only 35S label was associated with component C. The ratio of incorporated HS-CoM to component C was 1.96 to 1. The ratio of HS-CoM to factor F430, the nickel-containing cofactor of component C, was 1.18 to 1. Extraction of factor F430 from the protein resulted in the release of 62 +/- 8% of the 35S label, but the label was not covalently bound to F430. The incorporation of label into component C was coupled to methyl group reduction; no label was found associated with component C from a reconstituted reaction containing unlabeled 2-methylthioethanesulfonic acid and [thio-35S]HS-CoM.