BRENDA - Enzyme Database

Methyl-coenzyme M formation in methanogenic archaea. Involvement of zinc in coenzyme M activation

Sauer, K.; Thauer, R.K.; Eur. J. Biochem. 267, 2498-2504 (2000)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.1.1.246
overexpression of His-tagged inactive MtaA apoprotein in Escherichia coli strain M15 grown in the presence of 2 mM EDTA
Methanosarcina barkeri
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.1.1.246
EDTA
75% inhibition at 1 mM, complete inhibition at 2 mM, reversible by Zn2+ addition, competitive versus Zn2+, kinetics, overview
Methanosarcina barkeri
2.1.1.246
additional information
1 mm nitrilotriacetic acid has almost no effect on the MtaA activity
Methanosarcina barkeri
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.1.1.246
Co2+
required, the apoprotein reacts with zinc or cobalt to the fully active holoenzyme
Methanosarcina barkeri
2.1.1.246
additional information
Zn21 or Co21 are required for MtaA activity, Zn2+ can be replaced by Co2+ but not by Mg2+, the kinetics of activation by Co2+ being similarily slow. About 1 mol of transition metal is bound per mol of protein. The role of the transition metal in MtaA is to lower the microscopic pKa of the thiol group of coenzyme M by coordination to the zinc, and thus to increase its nucleophilicity for methyl group attack, pKZn2+ of MtaA is over 15
Methanosarcina barkeri
2.1.1.246
Zn2+
1 mol per mol of enzyme, required, the apoprotein reacts with zinc or cobalt to the fully active holoenzyme
Methanosarcina barkeri
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.1.1.246
50000
-
x * 50000
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]
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.1.1.246
Methanosarcina barkeri
-
gene mtaA
-
Purification (Commentary)
EC Number
Commentary
Organism
2.1.1.246
recombinant His-tagged apo MtaA from Escherichia coli strain M15 by nickel affinity and anionexchange chromatography
Methanosarcina barkeri
Reaction
EC Number
Reaction
Commentary
Organism
2.1.1.246
a [methyl-Co(III) methanol-specific corrinoid protein] + CoM = methyl-CoM + a [Co(I) methanol-specific corrinoid protein]
coenzyme M binds with its thiol group to the zinc in the active site of MtaA forming a coenzyme M thiolate zinc complex
Methanosarcina barkeri
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.1.1.246
2
-
purified Zn2+-containing holoenzyme, pH 7.0, 37°C
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] + coenzyme M
-
717562
Methanosarcina barkeri
methyl-CoM + a [Co(I) methanol-specific corrinoid protein]
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
2.1.1.246
?
x * 50000
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
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.1.246
7
-
assay at
Methanosarcina barkeri
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.246
overexpression of His-tagged inactive MtaA apoprotein in Escherichia coli strain M15 grown in the presence of 2 mM EDTA
Methanosarcina barkeri
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.1.1.246
EDTA
75% inhibition at 1 mM, complete inhibition at 2 mM, reversible by Zn2+ addition, competitive versus Zn2+, kinetics, overview
Methanosarcina barkeri
2.1.1.246
additional information
1 mm nitrilotriacetic acid has almost no effect on the MtaA activity
Methanosarcina barkeri
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.1.1.246
Co2+
required, the apoprotein reacts with zinc or cobalt to the fully active holoenzyme
Methanosarcina barkeri
2.1.1.246
additional information
Zn21 or Co21 are required for MtaA activity, Zn2+ can be replaced by Co2+ but not by Mg2+, the kinetics of activation by Co2+ being similarily slow. About 1 mol of transition metal is bound per mol of protein. The role of the transition metal in MtaA is to lower the microscopic pKa of the thiol group of coenzyme M by coordination to the zinc, and thus to increase its nucleophilicity for methyl group attack, pKZn2+ of MtaA is over 15
Methanosarcina barkeri
2.1.1.246
Zn2+
1 mol per mol of enzyme, required, the apoprotein reacts with zinc or cobalt to the fully active holoenzyme
Methanosarcina barkeri
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.1.1.246
50000
-
x * 50000
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]
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.246
recombinant His-tagged apo MtaA from Escherichia coli strain M15 by nickel affinity and anionexchange chromatography
Methanosarcina barkeri
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.1.1.246
2
-
purified Zn2+-containing holoenzyme, pH 7.0, 37°C
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] + coenzyme M
-
717562
Methanosarcina barkeri
methyl-CoM + a [Co(I) methanol-specific corrinoid protein]
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.1.1.246
?
x * 50000
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
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.1.246
7
-
assay at
Methanosarcina barkeri
General Information
EC Number
General Information
Commentary
Organism
2.1.1.246
additional information
MtaC and MtaB form a tight complex and the encoding genes form a transcription unit, whereas MtaA purifies separately and its encoding gene is located separately
Methanosarcina barkeri
2.1.1.246
physiological function
the methyltransferase designated MtaA together with the proteins MtaB and MtaC mediate the formation of methyl-coenzyme M from methanol and coenzyme M. MtaC is a 28-kDa corrinoid protein, MtaB, EC 2.1.1.90, catalyzes the methylation of MtaC and MtaA catalyzes the demethylation of methylated MtaC
Methanosarcina barkeri
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.1.1.246
additional information
MtaC and MtaB form a tight complex and the encoding genes form a transcription unit, whereas MtaA purifies separately and its encoding gene is located separately
Methanosarcina barkeri
2.1.1.246
physiological function
the methyltransferase designated MtaA together with the proteins MtaB and MtaC mediate the formation of methyl-coenzyme M from methanol and coenzyme M. MtaC is a 28-kDa corrinoid protein, MtaB, EC 2.1.1.90, catalyzes the methylation of MtaC and MtaA catalyzes the demethylation of methylated MtaC
Methanosarcina barkeri