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Literature summary extracted from

  • LeClerc, G.M.; Grahame, D.A.
    Methylcobamide:coenzyme M methyltransferase isozymes from Methanosarcina barkeri. Physicochemical characterization, cloning, sequence analysis, and heterologous gene expression (1996), J. Biol. Chem., 271, 18725-18731.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.246 genes cmtA and cmtM, DNA and amino acid sequence determination and analysis, expressionin Escherichia coli strain XL-1 Blue Methanosarcina barkeri
2.1.1.247 genes cmtA and cmtM, DNA and amino acid sequence determination and analysis, expression of the isozymes in Escherichia coli Methanosarcina barkeri

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.1.247 1,10-phenanthroline
-
Methanosarcina barkeri
2.1.1.247 EDTA
-
Methanosarcina barkeri
2.1.1.247 EGTA
-
Methanosarcina barkeri

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.246 additional information
-
additional information steady-state kinetics of isozymes MT2-A and MT2-M, overview Methanosarcina barkeri
2.1.1.247 additional information
-
additional information steady-state kinetics Methanosarcina barkeri
2.1.1.247 0.000014
-
[methyl-Co(III) trimethylamine-specific corrinoid protein] isozymes MT2-A and MT-2-M, pH 7.2, 23°C Methanosarcina barkeri
2.1.1.247 0.02
-
coenzyme M isozyme MT2-M, pH 7.2, 23°C Methanosarcina barkeri
2.1.1.247 0.035
-
coenzyme M isozyme MT2-A, pH 7.2, 23°C Methanosarcina barkeri
2.1.1.247 9
-
3-Mercaptopropionate isozyme MT2-A, pH 7.2, 23°C Methanosarcina barkeri
2.1.1.247 10
-
3-Mercaptopropionate isozyme MT2-M, pH 7.2, 23°C Methanosarcina barkeri

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.1.1.246 Zn2+
-
Methanosarcina barkeri
2.1.1.247 Zn2+ both isozymes are zinc-containing metalloproteins, zinc involvement in catalysis of coenzyme M methylation Methanosarcina barkeri

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.246 34000
-
x * 34000, isozymes MT2-A and MT2-M, SDS-PAGE, x * 36700, about, isozyme MT2-A, sequence calculation Methanosarcina barkeri
2.1.1.246 36700
-
x * 34000, isozymes MT2-A and MT2-M, SDS-PAGE, x * 36700, about, isozyme MT2-A, sequence calculation Methanosarcina barkeri

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.246 a [methyl-Co(III) methanol-specific corrinoid protein] + coenzyme M Methanosarcina barkeri
-
methyl-CoM + a [Co(I) methanol-specific corrinoid protein]
-
r
2.1.1.246 additional information Methanosarcina barkeri conversions of monomethylamine and dimethylamine to CH3-SCoM are dependent upon MT2-A, and are not supported by MT2-M. In contrast, MT2-M acts specifically in metabolism of methanol, but does not substitute for MT2-A in conversion of monomethylamine or dimethylamine. Nevertheless, both isozymes are capable of supporting the conversion of trimethylamine ?
-
?
2.1.1.247 HSCoM + methylcobalamin Methanosarcina barkeri
-
CH3-SCoM + cob(I)alamin + H+
-
r
2.1.1.247 [methyl-Co(III) trimethylamine-specific corrinoid protein] + coenzyme M Methanosarcina barkeri heterolytic cleavage of the methylcobamide carbon-cobalt bond with cob(I)alamin as the major product of the reaction methyl-CoM + [Co(I) trimethylamine-specific corrinoid protein]
-
r

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.246 Methanosarcina barkeri O30640 isozymes MT2-A and MT2-M encoded by genes cmtA and cmtM
-
2.1.1.247 Methanosarcina barkeri O30640 isozymes MT2-A and MT2-M encoded by genes cmtA and cmtM
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.1.1.246 cell culture the isozymes MT2-A and MT2-M are differentially expressed depending upon the substrate available for growth Methanosarcina barkeri
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.1.1.246 0.154
-
MT2 activity in recombinant Escherichia coli cells transfected with gene cmtM, pH 7.2, 37°C Methanosarcina barkeri
2.1.1.246 1.21
-
MT2 activity in recombinant Escherichia coli cells transfected with gene cmtA, pH 7.2, 37°C Methanosarcina barkeri
2.1.1.246 2.2
-
MT2 activity in cells grown on trimethylamine, pH 7.2, 37°C Methanosarcina barkeri
2.1.1.246 7.5
-
MT2 activity in cells grown on methanol, pH 7.2, 37°C Methanosarcina barkeri

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.246 a [methyl-Co(III) methanol-specific corrinoid protein] + 3-mercaptopropanoic acid
-
Methanosarcina barkeri methyl-3-mercaptopropanoic acid + a [Co(I) methanol-specific corrinoid protein]
-
r
2.1.1.246 a [methyl-Co(III) methanol-specific corrinoid protein] + coenzyme M
-
Methanosarcina barkeri methyl-CoM + a [Co(I) methanol-specific corrinoid protein]
-
r
2.1.1.246 a [methyl-Co(III) methanol-specific corrinoid protein] + coenzyme M the enzyme methylcobamide:CoM methyltransferase, MT2, catalyzes the transfer of the methyl group from the MT1-bound methylcobamide prosthetic group to coenzyme M Methanosarcina barkeri methyl-CoM + a [Co(I) methanol-specific corrinoid protein]
-
r
2.1.1.246 additional information conversions of monomethylamine and dimethylamine to CH3-SCoM are dependent upon MT2-A, and are not supported by MT2-M. In contrast, MT2-M acts specifically in metabolism of methanol, but does not substitute for MT2-A in conversion of monomethylamine or dimethylamine. Nevertheless, both isozymes are capable of supporting the conversion of trimethylamine Methanosarcina barkeri ?
-
?
2.1.1.247 HSCoM + methylcobalamin
-
Methanosarcina barkeri CH3-SCoM + cob(I)alamin + H+
-
r
2.1.1.247 additional information conversions of monomethylamine and dimethylamine to CH3-SCoM are dependent upon MT2-A, and are not supported by MT2-M, both isozymes catalyze S-methylation of 2-thioethanesulfonate, i.e. coenzyme M, and exhibit similar apparent Km values for coenzyme M, isozymes substrate specificities, overview Methanosarcina barkeri ?
-
?
2.1.1.247 [methyl-Co(III) dimethylamine-specific corrinoid protein] + coenzyme M isozyme MT2-A, not MT2-M Methanosarcina barkeri methyl-CoM + [Co(I) dimethylamine-specific corrinoid protein]
-
r
2.1.1.247 [methyl-Co(III) monomethylamine-specific corrinoid protein] + coenzyme M isozyme MT2-A, not MT2-M Methanosarcina barkeri methyl-CoM + [Co(I) monomethylamine-specific corrinoid protein]
-
r
2.1.1.247 [methyl-Co(III) trimethylamine-specific corrinoid protein] + 3-mercaptopropionate 3-mercaptopropionate is a coenzyme M analogue Methanosarcina barkeri methyl-3-mercaptopropionate + [Co(I) trimethylamine-specific corrinoid protein]
-
?
2.1.1.247 [methyl-Co(III) trimethylamine-specific corrinoid protein] + coenzyme M heterolytic cleavage of the methylcobamide carbon-cobalt bond with cob(I)alamin as the major product of the reaction Methanosarcina barkeri methyl-CoM + [Co(I) trimethylamine-specific corrinoid protein]
-
r
2.1.1.247 [methyl-Co(III) trimethylamine-specific corrinoid protein] + coenzyme M isozymes MT2-A and MT2-M, heterolytic cleavage of the methylcobamide carbon-cobalt bond with cob(I)alamin as the major product of the reaction Methanosarcina barkeri methyl-CoM + [Co(I) trimethylamine-specific corrinoid protein]
-
r

Subunits

EC Number Subunits Comment Organism
2.1.1.246 ? x * 34000, isozymes MT2-A and MT2-M, SDS-PAGE, x * 36700, about, isozyme MT2-A, sequence calculation Methanosarcina barkeri
2.1.1.246 More the isozymes MT2-A and MT2-M differ in their overall charge Methanosarcina barkeri
2.1.1.247 ? x * 36000-37000, recombinant isozymes MT2-A and MT2-M, SDS-PAGE Methanosarcina barkeri

Synonyms

EC Number Synonyms Comment Organism
2.1.1.246 methylcobamide:coenzyme M methyltransferase
-
Methanosarcina barkeri
2.1.1.246 methylcobamide:CoM methyltransferase
-
Methanosarcina barkeri
2.1.1.246 MT2-A
-
Methanosarcina barkeri
2.1.1.246 MT2-M
-
Methanosarcina barkeri
2.1.1.247 methylcobamide:coenzyme M methyltransferase
-
Methanosarcina barkeri
2.1.1.247 MT2
-
Methanosarcina barkeri

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.246 37
-
assay at Methanosarcina barkeri
2.1.1.247 23
-
assay at Methanosarcina barkeri

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.246 7.2
-
assay at Methanosarcina barkeri
2.1.1.247 7.2
-
assay at Methanosarcina barkeri

Cofactor

EC Number Cofactor Comment Organism Structure
2.1.1.247 additional information no enzyme-bound organic or inorganic cofactors Methanosarcina barkeri

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.1.1.247 0.005
-
pH 7.2, 23°C Methanosarcina barkeri EDTA
2.1.1.247 0.05
-
pH 7.2, 23°C Methanosarcina barkeri EGTA
2.1.1.247 0.07
-
pH 7.2, 23°C Methanosarcina barkeri 1,10-phenanthroline

General Information

EC Number General Information Comment Organism
2.1.1.247 additional information the enzyme shows an active site geometry in which coenzyme M is bound both by S-coordination to zinc, and electrostatic interaction of the sulfonate with a cationic group on the enzyme Methanosarcina barkeri