Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli as maltose binding fusion protein, wild-type and selenomethionine enzyme variants | Sus scrofa |
Crystallization (Comment) | Organism |
---|---|
20 mg/ml purified recombinant wild-type and selenomethionine enzyme, complexed with Mn2+ and isocitrate, 4°C, hanging drop vapour diffusion method, for the wild-type enzyme: 0.002 ml of enzyme solution containing 0.1 M triethanolamine chloride, pH 7.7, 0.15 M Na2SO4, 8 mM DL-isocitrate, 4 mM MnSO4, plus equal volume of 20% PEG 6000, 3% glycerol, against 0.75 ml reservoir solution containing 0.1 M triethanolamine chloride, pH 7.7, 0.15 M Na2SO4, 8 mM DL-isocitrate, 4 mM MnSO4, and 40% w/v xylitol, for the selenomethionine enzyme: 0.002 ml of enzyme solution containing 0.1 M triethanolamine chloride, pH 7.7, 0.15 M Na2SO4, 8 mM DL-isocitrate, plus equal volume of 18% PEG 6000, 3% glycerol, against 0.75 ml reservoir solution containing 0.1 M triethanolamine chloride, pH 7.7, 0.15 M Na2SO4, and 18% PEG 6000, 7-10 days, X-ray diffraction structure determination and analysis at 2.7 A resolution, modeling | Sus scrofa |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Sus scrofa | 5739 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | required | Sus scrofa |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
47000 | - |
2 * 47000, SDS-PAGE | Sus scrofa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NADP+ | Sus scrofa | - |
2-oxoglutarate + CO2 + NADPH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sus scrofa | P33198 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme variants fused to the maltose binding fusion protein, from Escherichia coli, to homogeneity by amylose affinity, and ion exchange chromatography | Sus scrofa |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
isocitrate + NADP+ = 2-oxoglutarate + CO2 + NADPH + H+ | active site structure contains a well-ordered Mn2+-isocitrate complex, reaction mechanism | Sus scrofa |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
heart | - |
Sus scrofa | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NADP+ | - |
Sus scrofa | 2-oxoglutarate + CO2 + NADPH + H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 47000, SDS-PAGE | Sus scrofa |
Synonyms | Comment | Organism |
---|---|---|
NADP+-dependent isocitrate dehydrogenase | - |
Sus scrofa |
NADP-IDH | - |
Sus scrofa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | dependent on | Sus scrofa |