BRENDA - Enzyme Database

Methylcobamide:coenzyme M methyltransferase isozymes from Methanosarcina barkeri. Physicochemical characterization, cloning, sequence analysis, and heterologous gene expression

LeClerc, G.M.; Grahame, D.A.; J. Biol. Chem. 271, 18725-18731 (1996)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
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
Commentary
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
Commentary
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, 23C
Methanosarcina barkeri
2.1.1.247
0.02
-
coenzyme M
isozyme MT2-M, pH 7.2, 23C
Methanosarcina barkeri
2.1.1.247
0.035
-
coenzyme M
isozyme MT2-A, pH 7.2, 23C
Methanosarcina barkeri
2.1.1.247
9
-
3-Mercaptopropionate
isozyme MT2-A, pH 7.2, 23C
Methanosarcina barkeri
2.1.1.247
10
-
3-Mercaptopropionate
isozyme MT2-M, pH 7.2, 23C
Methanosarcina barkeri
Metals/Ions
EC Number
Metals/Ions
Commentary
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]
Commentary
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
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
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
Primary Accession No. (UniProt)
Commentary
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
Commentary
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 [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
2.1.1.246
0.154
-
MT2 activity in recombinant Escherichia coli cells transfected with gene cmtM, pH 7.2, 37C
Methanosarcina barkeri
2.1.1.246
1.21
-
MT2 activity in recombinant Escherichia coli cells transfected with gene cmtA, pH 7.2, 37C
Methanosarcina barkeri
2.1.1.246
2.2
-
MT2 activity in cells grown on trimethylamine, pH 7.2, 37C
Methanosarcina barkeri
2.1.1.246
7.5
-
MT2 activity in cells grown on methanol, pH 7.2, 37C
Methanosarcina barkeri
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.246
a [methyl-Co(III) methanol-specific corrinoid protein] + 3-mercaptopropanoic acid
-
717753
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
-
717753
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
717753
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
717753
Methanosarcina barkeri
?
-
-
-
-
2.1.1.247
HSCoM + methylcobalamin
-
717753
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
717753
Methanosarcina barkeri
?
-
-
-
-
2.1.1.247
[methyl-Co(III) dimethylamine-specific corrinoid protein] + coenzyme M
isozyme MT2-A, not MT2-M
717753
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
717753
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
717753
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
717753
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
717753
Methanosarcina barkeri
methyl-CoM + [Co(I) trimethylamine-specific corrinoid protein]
-
-
-
r
Subunits
EC Number
Subunits
Commentary
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
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
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
Commentary
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
Commentary
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
Commentary
Organism
Inhibitor
Structure
2.1.1.247
0.005
-
pH 7.2, 23C
Methanosarcina barkeri
EDTA
2.1.1.247
0.05
-
pH 7.2, 23C
Methanosarcina barkeri
EGTA
2.1.1.247
0.07
-
pH 7.2, 23C
Methanosarcina barkeri
1,10-phenanthroline
Cloned(Commentary) (protein specific)
EC Number
Commentary
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
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.1.1.247
additional information
no enzyme-bound organic or inorganic cofactors
Methanosarcina barkeri
IC50 Value (protein specific)
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
2.1.1.247
0.005
-
pH 7.2, 23C
Methanosarcina barkeri
EDTA
2.1.1.247
0.05
-
pH 7.2, 23C
Methanosarcina barkeri
EGTA
2.1.1.247
0.07
-
pH 7.2, 23C
Methanosarcina barkeri
1,10-phenanthroline
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
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] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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, 23C
Methanosarcina barkeri
2.1.1.247
0.02
-
coenzyme M
isozyme MT2-M, pH 7.2, 23C
Methanosarcina barkeri
2.1.1.247
0.035
-
coenzyme M
isozyme MT2-A, pH 7.2, 23C
Methanosarcina barkeri
2.1.1.247
9
-
3-Mercaptopropionate
isozyme MT2-A, pH 7.2, 23C
Methanosarcina barkeri
2.1.1.247
10
-
3-Mercaptopropionate
isozyme MT2-M, pH 7.2, 23C
Methanosarcina barkeri
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
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] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
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) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
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
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
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] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
2.1.1.246
0.154
-
MT2 activity in recombinant Escherichia coli cells transfected with gene cmtM, pH 7.2, 37C
Methanosarcina barkeri
2.1.1.246
1.21
-
MT2 activity in recombinant Escherichia coli cells transfected with gene cmtA, pH 7.2, 37C
Methanosarcina barkeri
2.1.1.246
2.2
-
MT2 activity in cells grown on trimethylamine, pH 7.2, 37C
Methanosarcina barkeri
2.1.1.246
7.5
-
MT2 activity in cells grown on methanol, pH 7.2, 37C
Methanosarcina barkeri
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.246
a [methyl-Co(III) methanol-specific corrinoid protein] + 3-mercaptopropanoic acid
-
717753
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
-
717753
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
717753
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
717753
Methanosarcina barkeri
?
-
-
-
-
2.1.1.247
HSCoM + methylcobalamin
-
717753
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
717753
Methanosarcina barkeri
?
-
-
-
-
2.1.1.247
[methyl-Co(III) dimethylamine-specific corrinoid protein] + coenzyme M
isozyme MT2-A, not MT2-M
717753
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
717753
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
717753
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
717753
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
717753
Methanosarcina barkeri
methyl-CoM + [Co(I) trimethylamine-specific corrinoid protein]
-
-
-
r
Subunits (protein specific)
EC Number
Subunits
Commentary
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
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
2.1.1.246
37
-
assay at
Methanosarcina barkeri
2.1.1.247
23
-
assay at
Methanosarcina barkeri
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.1.246
7.2
-
assay at
Methanosarcina barkeri
2.1.1.247
7.2
-
assay at
Methanosarcina barkeri
General Information
EC Number
General Information
Commentary
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
General Information (protein specific)
EC Number
General Information
Commentary
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