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J Bacteriol. 1995 August; 177(15): 4410–4416.
PMCID: PMC177191

Involvement of the "A" isozyme of methyltransferase II and the 29-kilodalton corrinoid protein in methanogenesis from monomethylamine.


An assay which allowed detection of proteins involved in the trimethylamine- or monomethylamine (MMA)-dependent methylation of coenzyme M (CoM) was developed. The two activities could be separated by anion-exchange chromatography. The unresolved activity responsible for MMA:CoM methyl transfer eluted from a gel permeation column in the molecular mass range of 32 kDa. The activity was purified to two monomeric proteins of 40 and 29 kDa. The preparation contained protein-bound corrinoid in a mixture of Co(II) and Co(III) states, as well as methyl-B12:CoM methyltransferase (MT2) activity. N-terminal sequence analysis demonstrated that the polypeptides were two previously identified proteins of undefined physiological function. The smaller polypeptide was the monomeric 29-kDa corrinoid protein. The larger polypeptide was the "A" isozyme of MT2. Individually purified preparations of both proteins increased the rate of MMA-dependent CoM methylation by approximately 1.7 mumol/min/mg of purified protein above background activity in the extract of methanol-grown cells. These results indicate that the 29-kDa corrinoid protein and the "A" isozyme of MT2 function in methanogenesis from MMA. A likely mechanism is that the 29-kDa corrinoid is methylated by MMA and the methyl group is then transferred by the "A" isozyme of MT2 to CoM.

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Selected References

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  • Banerjee RV, Harder SR, Ragsdale SW, Matthews RG. Mechanism of reductive activation of cobalamin-dependent methionine synthase: an electron paramagnetic resonance spectroelectrochemical study. Biochemistry. 1990 Feb 6;29(5):1129–1135. [PubMed]
  • Becher B, Müller V, Gottschalk G. N5-methyl-tetrahydromethanopterin:coenzyme M methyltransferase of Methanosarcina strain Gö1 is an Na(+)-translocating membrane protein. J Bacteriol. 1992 Dec;174(23):7656–7660. [PMC free article] [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Cao XJ, Krzycki JA. Acetate-dependent methylation of two corrinoid proteins in extracts of Methanosarcina barkeri. J Bacteriol. 1991 Sep;173(17):5439–5448. [PMC free article] [PubMed]
  • Daas PJ, Gerrits KA, Keltjens JT, van der Drift C, Vogels GD. Involvement of an activation protein in the methanol:2-mercaptoethanesulfonic acid methyltransferase reaction in Methanosarcina barkeri. J Bacteriol. 1993 Mar;175(5):1278–1283. [PMC free article] [PubMed]
  • ELLMAN GL. A colorimetric method for determining low concentrations of mercaptans. Arch Biochem Biophys. 1958 Apr;74(2):443–450. [PubMed]
  • Ferry JG. Biochemistry of methanogenesis. Crit Rev Biochem Mol Biol. 1992;27(6):473–503. [PubMed]
  • Ferry JG. Methane from acetate. J Bacteriol. 1992 Sep;174(17):5489–5495. [PMC free article] [PubMed]
  • Fischer R, Gärtner P, Yeliseev A, Thauer RK. N5-methyltetrahydromethanopterin: coenzyme M methyltransferase in methanogenic archaebacteria is a membrane protein. Arch Microbiol. 1992;158(3):208–217. [PubMed]
  • Gärtner P, Ecker A, Fischer R, Linder D, Fuchs G, Thauer RK. Purification and properties of N5-methyltetrahydromethanopterin:coenzyme M methyltransferase from Methanobacterium thermoautotrophicum. Eur J Biochem. 1993 Apr 1;213(1):537–545. [PubMed]
  • Grahame DA. Different isozymes of methylcobalamin:2-mercaptoethanesulfonate methyltransferase predominate in methanol- versus acetate-grown Methanosarcina barkeri. J Biol Chem. 1989 Aug 5;264(22):12890–12894. [PubMed]
  • Grahame DA. Catalysis of acetyl-CoA cleavage and tetrahydrosarcinapterin methylation by a carbon monoxide dehydrogenase-corrinoid enzyme complex. J Biol Chem. 1991 Nov 25;266(33):22227–22233. [PubMed]
  • Grahame DA. Substrate and cofactor reactivity of a carbon monoxide dehydrogenase-corrinoid enzyme complex: stepwise reduction of iron-sulfur and corrinoid centers, the corrinoid Co2+/1+ redox midpoint potential, and overall synthesis of acetyl-CoA. Biochemistry. 1993 Oct 12;32(40):10786–10793. [PubMed]
  • Harder SR, Lu WP, Feinberg BA, Ragsdale SW. Spectroelectrochemical studies of the corrinoid/iron-sulfur protein involved in acetyl coenzyme A synthesis by Clostridium thermoaceticum. Biochemistry. 1989 Nov 14;28(23):9080–9087. [PubMed]
  • Hippe H, Caspari D, Fiebig K, Gottschalk G. Utilization of trimethylamine and other N-methyl compounds for growth and methane formation by Methanosarcina barkeri. Proc Natl Acad Sci U S A. 1979 Jan;76(1):494–498. [PubMed]
  • Jablonski PE, Lu WP, Ragsdale SW, Ferry JG. Characterization of the metal centers of the corrinoid/iron-sulfur component of the CO dehydrogenase enzyme complex from Methanosarcina thermophila by EPR spectroscopy and spectroelectrochemistry. J Biol Chem. 1993 Jan 5;268(1):325–329. [PubMed]
  • Kengen SW, Daas PJ, Duits EF, Keltjens JT, van der Drift C, Vogels GD. Isolation of a 5-hydroxybenzimidazolyl cobamide-containing enzyme involved in the methyltetrahydromethanopterin: coenzyme M methyltransferase reaction in Methanobacterium thermoautotrophicum. Biochim Biophys Acta. 1992 Feb 1;1118(3):249–260. [PubMed]
  • Kiene RP, Oremland RS, Catena A, Miller LG, Capone DG. Metabolism of reduced methylated sulfur compounds in anaerobic sediments and by a pure culture of an estuarine methanogen. Appl Environ Microbiol. 1986 Nov;52(5):1037–1045. [PMC free article] [PubMed]
  • Kremer JD, Cao X, Krzycki J. Isolation of two novel corrinoid proteins from acetate-grown Methanosarcina barkeri. J Bacteriol. 1993 Aug;175(15):4824–4833. [PMC free article] [PubMed]
  • Krzycki JA, Mortenson LE, Prince RC. Paramagnetic centers of carbon monoxide dehydrogenase from aceticlastic Methanosarcina barkeri. J Biol Chem. 1989 May 5;264(13):7217–7221. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Oremland RS, Kiene RP, Mathrani I, Whiticar MJ, Boone DR. Description of an estuarine methylotrophic methanogen which grows on dimethyl sulfide. Appl Environ Microbiol. 1989 Apr;55(4):994–1002. [PMC free article] [PubMed]
  • Pol A, van der Drift C, Vogels GD. Corrinoids from Methanosarcina barkeri: structure of the alpha-ligand. Biochem Biophys Res Commun. 1982 Sep 30;108(2):731–737. [PubMed]
  • Stadtman TC, Blaylock BA. Role of B12 compounds in methane formation. Fed Proc. 1966 Nov-Dec;25(6):1657–1661. [PubMed]
  • Stupperich E, Eisinger HJ, Albracht SP. Evidence for a super-reduced cobamide as the major corrinoid fraction in vivo and a histidine residue as a cobalt ligand of the p-cresolyl cobamide in the acetogenic bacterium Sporomusa ovata. Eur J Biochem. 1990 Oct 5;193(1):105–109. [PubMed]
  • Taylor CD, Wolfe RS. Structure and methylation of coenzyme M(HSCH2CH2SO3). J Biol Chem. 1974 Aug 10;249(15):4879–4885. [PubMed]
  • Terlesky KC, Nelson MJ, Ferry JG. Isolation of an enzyme complex with carbon monoxide dehydrogenase activity containing corrinoid and nickel from acetate-grown Methanosarcina thermophila. J Bacteriol. 1986 Dec;168(3):1053–1058. [PMC free article] [PubMed]
  • van der Maarel M, Jansen M, Hansen TA. Methanogenic conversion of 3-s-methylmercaptopropionate to 3-mercaptopropionate. Appl Environ Microbiol. 1995 Jan;61(1):48–51. [PMC free article] [PubMed]
  • van der Meijden P, Heythuysen HJ, Pouwels A, Houwen F, van der Drift C, Vogels GD. Methyltransferases involved in methanol conversion by Methanosarcina barkeri. Arch Microbiol. 1983 Jun;134(3):238–242. [PubMed]
  • van der Meijden P, te Brömmelstroet BW, Poirot CM, van der Drift C, Vogels GD. Purification and properties of methanol:5-hydroxybenzimidazolylcobamide methyltransferase from Methanosarcina barkeri. J Bacteriol. 1984 Nov;160(2):629–635. [PMC free article] [PubMed]
  • Walther R, Fahlbusch K, Sievert R, Gottschalk G. Formation of trideuteromethane from deuterated trimethylamine or methylamine by Methanosarcina barkeri. J Bacteriol. 1981 Oct;148(1):371–373. [PMC free article] [PubMed]
  • Weimer PJ, Zeikus JG. One carbon metabolism in methanogenic bacteria. Cellular characterization and growth of Methanosarcina barkeri. Arch Microbiol. 1978 Oct 4;119(1):49–57. [PubMed]
  • Yeliseev A, Gärtner P, Harms U, Linder D, Thauer RK. Function of methylcobalamin: coenzyme M methyltransferase isoenzyme II in Methanosarcina barkeri. Arch Microbiol. 1993;159(6):530–536. [PubMed]

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