Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta(DE3)pLysS cells | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
E392A | site-directed mutagenesis, kcat is reduced 12fold compared to the wild-type enzyme. The mutant shows no detectable adenosylcobalamin homolysis upon binding of the physiological substrate | Homo sapiens |
E392D | site-directed mutagenesis, kcat is reduced 330fold compared to the wild-type enzyme. The mutant shows no detectable adenosylcobalamin homolysis upon binding of the physiological substrate | Homo sapiens |
E392Q | site-directed mutagenesis, kcat is reduced 16fold compared to the wild-type enzyme. The mutant shows no detectable adenosylcobalamin homolysis upon binding of the physiological substrate | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | pre-steady-state and steady-state kinetics of wild-type and mutant enzymes | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Homo sapiens | 5739 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-methylmalonyl-CoA | Homo sapiens | - |
succinyl-CoA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P22033 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta(DE3)pLysS cells by nickel affinity and anion exchange chromatography | Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(R)-methylmalonyl-CoA = succinyl-CoA | mechanism, overview, a gradual weakening of the electrostatic energy between the protein and the ribose leads to a progressive increase in the activation energy barrier for Co-C bond homolysis, key role for the conserved polar glutamate residue in controlling the initial generation of radical species | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-methylmalonyl-CoA | - |
Homo sapiens | succinyl-CoA | - |
? | |
(R)-methylmalonyl-CoA | binding of substrate to the enzyme leads to the formation of an electrostatic interaction between a conserved glutamate side chain and the adenosyl ribose of the adenosylcobalamin cofactor. Residue Glu392 is involved in adenosylcobalamin Co-C bond labilization and catalysis | Homo sapiens | succinyl-CoA | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MCM | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.012 | - |
(R)-methylmalonyl-CoA | recombinant His-tagged mutant E392D, pH 8.5, 25°C | Homo sapiens | |
0.25 | - |
(R)-methylmalonyl-CoA | recombinant His-tagged mutant E392Q, pH 8.5, 25°C | Homo sapiens | |
0.32 | - |
(R)-methylmalonyl-CoA | recombinant His-tagged mutant E392A, pH 8.5, 25°C | Homo sapiens | |
3.95 | - |
(R)-methylmalonyl-CoA | recombinant His-tagged wild-type enzyme, pH 8.5, 25°C | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
adenosylcobalamin | binding analysis with recombinant wild-type and mutant enzymes, overview | Homo sapiens |