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Literature summary for 1.1.1.42 extracted from
- Evaluation by mutagenesis of the roles of His309, His315, and His319 in the coenzyme site of pig heart NADP-dependent isocitrate dehydrogenase (2002), Biochemistry, 41, 5637-5643.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of wild-type and mutant enzymes in Escherichia coli as maltose binding fusion proteins | Sus scrofa |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| H309F | site-directed mutagenesis, inactive mutant, poor cofactor binding, altered secondary structure | Sus scrofa |
| H309Q | site-directed mutagenesis, inactive mutant, poor cofactor binding, altered secondary structure | Sus scrofa |
| H315Q | site-directed mutagenesis, 40fold increased Km for NADP+ compared to the wild-type enzyme | Sus scrofa |
| H319Q | site-directed mutagenesis, cofactor binding and kinetics similar to the wild-type enzyme, slightly reduced activity | Sus scrofa |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| NADPH | - |
Sus scrofa |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | measurement methods, the maltose binding fusion protein of the recombinant enzymes alters the kinetic parameters, overview | Sus scrofa | |
| 0.00033 | - |
Mn2+ | pH 7.4, 25°C, recombinant wild-type | Sus scrofa | |
| 0.00039 | - |
Mn2+ | pH 7.4, 25°C, recombinant mutant H319Q | Sus scrofa | |
| 0.0027 | - |
Mn2+ | pH 7.4, 25°C, recombinant mutant H315Q | Sus scrofa | |
| 0.0051 | - |
NADP+ | pH 7.4, 25°C, recombinant mutant H319Q | Sus scrofa | |
| 0.0056 | - |
NADP+ | pH 7.4, 25°C, recombinant wild-type | Sus scrofa | |
| 0.0083 | - |
DL-isocitrate | pH 7.4, 25°C, recombinant mutant H319Q | Sus scrofa | |
| 0.0084 | - |
DL-isocitrate | pH 7.4, 25°C, recombinant wild-type | Sus scrofa | |
| 0.0087 | - |
DL-isocitrate | pH 7.4, 25°C, recombinant mutant H315Q | Sus scrofa | |
| 0.218 | - |
NADP+ | pH 7.4, 25°C, recombinant mutant H315Q | Sus scrofa | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| mitochondrion | - |
Sus scrofa | 5739 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mn2+ | required, not bound normally | Sus scrofa | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 46600 | - |
2 * 46600, recombinant thrombin cleaved wild-type and mutant enzymes, SDS-PAGE | Sus scrofa |
| 90000 | - |
recombinant maltose binding fusion proteins, wild-type and mutant enzymes, gel filtration | Sus scrofa |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| DL-isocitrate + NADP+ | Sus scrofa | - |
2-oxoglutarate + CO2 + NADPH | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Sus scrofa | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and mutant maltose binding fusion proteins from Escherichia coli by amylose affinity and size exclusion chromatography | Sus scrofa |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| heart | - |
Sus scrofa | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 4.1 | - |
purified recombinant mutant H315Q | Sus scrofa |
| 20.1 | - |
purified recombinant mutant H319Q | Sus scrofa |
| 37.8 | - |
purified recombinant wild-type enzyme | Sus scrofa |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| DL-isocitrate + NADP+ | - |
Sus scrofa | 2-oxoglutarate + CO2 + NADPH | - |
? | |
| Ds-isocitrate + NADP+ | - |
Sus scrofa | 2-oxoglutarate + CO2 + NADPH | - |
? | |
| additional information | His319 and His315 are not responsible for enzyme Fe2+-isocitrate cleavage | Sus scrofa | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 46600, recombinant thrombin cleaved wild-type and mutant enzymes, SDS-PAGE | Sus scrofa |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| NADP-dependent isocitrate dehydrogenase | - |
Sus scrofa |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Sus scrofa |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.4 | - |
assay at | Sus scrofa |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5 | 8 | - |
Sus scrofa |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADP+ | binding of 1 mol NADP+ per enzyme subunit, His309 is highly important for cofactor binding, HIs315 is involved, while His319 is not | Sus scrofa | |
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