Chloromethane: tetrahydrofolate methyl transfer by two proteins from Methylobacterium chloromethanicum strain CM4.
The cmuA and cmuB genes are required for growth of Methylobacterium chloromethanicum strain CM4 with chloromethane as the sole carbon source. While CmuB was previously shown to possess methylcobalamin:tetrahydrofolate methyltransferase activity, sequence analysis indicated that CmuA represented a novel and so far unique two-domain methyltransferase/corrinoid-binding protein involved in methyl transfer from chloromethane to a corrin moiety. CmuA was purified from wild-type M. chloromethanicum strain CM4 and characterized as a monomeric, cobalt-containing and zinc-containing enzyme of molecular mass 67 kDa with a bound vitamin B12 cofactor. In combination, CmuA and CmuB proteins catalyze the in vitro transfer of the methyl group of chloromethane to tetrahydrofolate, thus affording a direct link between chloromethane dehalogenation and core C1 metabolism of Methylobacterium. Chloromethane dehalogenase activity in vitro is limited by CmuB, as formation of methyltetrahydrofolate from chloromethane displays apparent Michaelis-Menten kinetics with respect to methylated CmuA, with an apparent Km of 0.27 microM and a Vmax of 0.45 U x mg(-1). This contrasts with sequence-related systems for methyl transfer from methanogens, which involve methyltransferase and corrinoid protein components in well-defined stoichiometric ratios.
Studer
A
A
Institut für Mikrobiologie, ETH-Zentrum, Zürich, Switzerland.
Stupperich
E
E
Vuilleumier
S
S
Leisinger
T
T
D001426:Bacterial Proteins; D002244:Carbon / Q000378:metabolism; D002384:Catalysis; D002474:Cell-Free System; D003035:Cobalt / Q000737:chemistry / Q000494:pharmacology; D005492:Folic Acid / Q000737:chemistry; D007700:Kinetics; D008190:Lyases / Q000737:chemistry* / Q000378:metabolism; D008737:Methyl Chloride / Q000737:chemistry*; D020580:Methylobacterium / Q000737:chemistry*; D008780:Methyltransferases / Q000737:chemistry* / Q000302:isolation & purification; D008954:Models, Biological; D017434:Protein Structure, Tertiary; D013763:Tetrahydrofolates / Q000737:chemistry*; D013997:Time Factors
D001426:Bacterial Proteins; D013763:Tetrahydrofolates; D003035:Cobalt; C030371:5,6,7,8-tetrahydrofolic acid; D002244:Carbon; D005492:Folic Acid; D008737:Methyl Chloride; C118740:CmuA protein, Methylobacterium chloromethanicum; D008780:Methyltransferases; D008190:Lyases; C045224:dichloromethane dehalogenase