Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, expression of wild-type and mutant enzymes in Escherichia coli strain JM109 | Columba sp. |
Protein Variants | Comment | Organism |
---|---|---|
K162A | site-directed mutagenesis, the mutation does not affect Mn2+ binding of the mutant enzyme, but kcat is 27000fold reduced compared to the wild-type enzyme, NH4Cl shows no rescue of the pyruvate reduction in the K162A mutant, while for oxaloacetate decarboxylation, ammonium chloride demonstrated a maximum restoration of 3.5fold at 1 mM, and its rescue efficiency decreases with increasing concentration | Columba sp. |
K162Q | site-directed mutagenesis, the mutation does not affect Mn2+ binding of the mutant enzyme, but kcat is 3500fold reduced compared to the wild-type enzyme | Columba sp. |
K162R | site-directed mutagenesis,the mutation does not affect Mn2+ binding of the mutant enzyme, but kcat is 125fold reduced compared to the wild-type enzyme | Columba sp. |
Y91F | site-directed mutagenesis, the mutation does not affect Mn2+ binding of the mutant enzyme, the mutant shows a 25fold increase and a 3fold decrease in the Km values for (S)-malate and NADP+ respectively, and its kcat value is decreased by 200fold compared to wild-type enzyme | Columba sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(S)-malate | substrate inhibition | Columba sp. | |
NADP+ | substrate inhibition | Columba sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of mutant enzymes, overview | Columba sp. | |
0.0034 | - |
NADP+ | pH 4.5, 25°C, wild-type enzyme | Columba sp. | |
0.23 | - |
(S)-malate | pH 4.5, 25°C, wild-type enzyme | Columba sp. |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Columba sp. | 5829 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | required for forward and reverse reaction, residues Tyr91 and Lys162 are not involved in metal ion binding | Columba sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NADP+ | Columba sp. | - |
pyruvate + CO2 + NADPH | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Columba sp. | - |
gene maeB encoding isozyme MaeB | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain JM109 by anion exchange chromatography and 2',5'-ADP affinity chromatography to over 95% purity | Columba sp. |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(S)-malate + NADP+ = pyruvate + CO2 + NADPH + H+ | catalytic mechanism | Columba sp. |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
liver | - |
Columba sp. | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NADP+ | - |
Columba sp. | pyruvate + CO2 + NADPH | - |
r | |
(S)-malate + NADP+ | via oxaloacetate, roles of Tyr91 and Lys162 in general acid-base catalysis in the pigeon NADP+-dependent malic enzyme, overview | Columba sp. | pyruvate + CO2 + NADPH | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | circular dichroism structure analysis of wild-type and mutant enzymes, overview | Columba sp. |
Synonyms | Comment | Organism |
---|---|---|
MaeB | - |
Columba sp. |
NADP+-dependent malic enzyme | - |
Columba sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Columba sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
140.1 | - |
(S)-malate | pH 4.5, 25°C, wild-type enzyme | Columba sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.5 | - |
assay at, forward reaction | Columba sp. |
7.4 | - |
assay at, reverse reaction | Columba sp. |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
pH profiles of wild-type and mutant enzymes, overview | Columba sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Columba sp. | |
NADPH | - |
Columba sp. |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0089 | - |
NADP+ | pH 4.5, 25°C, wild-type enzyme | Columba sp. | |
0.28 | - |
(S)-malate | pH 4.5, 25°C, wild-type enzyme | Columba sp. |