Activating Compound | Comment | Organism | Structure |
---|---|---|---|
AMP | allosteric activation, isocitrate binding is essential for binding of AMP to IDH1 subunits | Saccharomyces cerevisiae | |
AMP | four AMP binding sites located on subunit IDH2 | Saccharomyces cerevisiae | |
DTT | - |
Saccharomyces cerevisiae | |
DTT | decreases disulfide content of the enzyme, kinetics of enzyme mutants in presence or absence of DTT, overview | Saccharomyces cerevisiae |
Cloned (Comment) | Organism |
---|---|
His-tagged enzyme expression in Escherichia coli strain BL21-Gold(DE3) | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
C150S | an octameric IDH1/IDH2C150S mutant enzyme, that shows unaltered activity and similar kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
C150S | site-directed mutagenesis, IDH1/IDH2C150S octameric enzyme | Saccharomyces cerevisiae |
C56S/C150S/C242S | site-directed mutagenesis, IDH1/IDH2C150S octameric enzyme | Saccharomyces cerevisiae |
C56S/C242S | an octameric IDH1/IDH2C56S/C242S mutant enzyme, the mutant enzyme shows a reduction in Vmax relative to that of the wild-type enzyme of about 50%, although about 30% activity is restored in the presence of dithiothreitol | Saccharomyces cerevisiae |
C56S/C242S | site-directed mutagenesis, IDH1/IDH2C150S octameric enzyme | Saccharomyces cerevisiae |
G15D | a tetrameric IDH1G15D/IDH2 mutant enzyme, that shows reduced activity and altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Diamide | increases disulfide content of the enzyme | Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | NAD+-specific D-isocitrate dehydrogenase is an allosterically regulated. AMP and NAD+ binding kinetics of enzyme mutants in presence or absence of DTT, overview | Saccharomyces cerevisiae | |
additional information | - |
additional information | yeast NAD+-specific isocitrate dehydrogenase is an allosterically regulated | Saccharomyces cerevisiae | |
0.09 | - |
D-isocitrate | octameric wild-type enzyme with AMP, without DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.1 | - |
D-isocitrate | octameric enzyme mutant IDH1/IDH2C150S with AMP, with or without DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.1 | - |
D-isocitrate | octameric wild-type enzyme with AMP and DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.17 | - |
D-isocitrate | octameric enzyme mutant IDH1/IDH2C56S/C150S/C242S with AMP, without DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.19 | - |
D-isocitrate | octameric enzyme mutant IDH1/IDH2C56S/C150S/C242S with AMP and DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.2 | - |
D-isocitrate | octameric enzyme mutant IDH1/IDH2C56S/C242S with AMP and DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.32 | - |
D-isocitrate | octameric enzyme mutant IDH1/IDH2C56S/C242S with AMP, without DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.48 | - |
D-isocitrate | octameric enzyme IDH1G15D/IDH2 with AMP and DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.5 | - |
D-isocitrate | octameric enzyme IDH1G15D/IDH2 with AMP, without DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.51 | - |
D-isocitrate | octameric wild-type enzyme without AMP, with DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.53 | - |
D-isocitrate | octameric enzyme mutant IDH1/IDH2C150S without AMP and DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.54 | - |
D-isocitrate | octameric enzyme mutant IDH1/IDH2C150S without AMP, with DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
0.56 | - |
D-isocitrate | octameric wild-type enzyme without AMP and DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
1.01 | - |
D-isocitrate | octameric enzyme mutant IDH1G15D/IDH2 without AMP and DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
1.03 | - |
D-isocitrate | octameric enzyme mutant IDH1/IDH2C56S/C150S/C242S without AMP, with or without DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
1.19 | - |
D-isocitrate | octameric enzyme IDH1G15D/IDH2 without AMP, with DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
1.3 | - |
D-isocitrate | octameric enzyme mutant IDH1/IDH2C56S/C242S without AMP, with DTT, pH 7.4, 24°C | Saccharomyces cerevisiae | |
1.6 | - |
D-isocitrate | octameric enzyme mutant IDH1/IDH2C56S/C242S without AMP and DTT, pH 7.4, 24°C | Saccharomyces cerevisiae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required, activates | Saccharomyces cerevisiae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
37755 | - |
4 * 38001, subunit IDH1, + 4 * 37755, subunit IDH2, sequence calculation | Saccharomyces cerevisiae |
38001 | - |
4 * 38001, subunit IDH1, + 4 * 37755, subunit IDH2, sequence calculation | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-isocitrate + NAD+ | Saccharomyces cerevisiae | - |
2-oxoglutarate + CO2 + NADH | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21-Gold(DE3) by nickel affinity chromatography | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-isocitrate + NAD+ | - |
Saccharomyces cerevisiae | 2-oxoglutarate + CO2 + NADH | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme is composed of four heterodimers, in two heterotetramers, of a catalytic IDH2 subunit and a regulatory IDH1 subunit, IDH octamer structure, overview | Saccharomyces cerevisiae |
octamer | 4 * 38001, subunit IDH1, + 4 * 37755, subunit IDH2, sequence calculation | Saccharomyces cerevisiae |
octamer | the enzyme is composed of four heterodimers of a catalytic IDH2 subunit and a regulatory IDH1 subunit | Saccharomyces cerevisiae |
Synonyms | Comment | Organism |
---|---|---|
NAD+-specific isocitrate dehydrogenase | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
24 | - |
assay at | Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Saccharomyces cerevisiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | dependent on, one NAD+ site per enzyme tetramer, two functional NAD+ binding sites in the wild-type enzyme | Saccharomyces cerevisiae | |
NAD+ | two NAD+ binding sites located on subunit IDH2 | Saccharomyces cerevisiae |
General Information | Comment | Organism |
---|---|---|
additional information | interactions between sulfhydryl side chains of IDH2 Cys150 residues limit access to eight D-isocitrate and four AMP binding sites. In the presence of dithiothreitol, all ligand binding sites except for two potential NAD+ sites can be occupied | Saccharomyces cerevisiae |
additional information | the IDH2 Cys-150 residue controls access to isocitrate binding sites. The wild-type enzyme displays four binding sites for isocitrate and two binding sites for AMP in the absence of dithiothreitol, and these numbers increase to eight and four in the presence of dithiothreitol, respectively | Saccharomyces cerevisiae |