Activating Compound | Comment | Organism | Structure |
---|---|---|---|
acetic acid | mutant Y140T, 106fold activation compared to the wild-type enzyme | Sus scrofa | |
ethylamine | mutant K212Q, 4fold activation compared to the wild-type enzyme | Sus scrofa | |
phenol | mutant Y140T, 88fold activation compared to the wild-type enzyme | Sus scrofa |
Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Sus scrofa |
Protein Variants | Comment | Organism |
---|---|---|
K212Q | site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, altered pH-dependency of the activity | Sus scrofa |
K212R | site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, altered pH-dependency of the activity | Sus scrofa |
K212Y | site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, altered pH-dependency of the activity | Sus scrofa |
Y140E | site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, unaltered Km for isocitrate and NADP+ | Sus scrofa |
Y140F | site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, unaltered Km for isocitrate and NADP+ | Sus scrofa |
Y140K | site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, unaltered Km for isocitrate and NADP+ | Sus scrofa |
Y140T | site-directed mutagenesis, highly decreased activity in both reaction directions compared to the wild-type enzyme, unaltered Km for isocitrate and NADP+, highly increased activation by added exogenous acetic acid and phenol compared to the wild-type enzyme | Sus scrofa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00004 | - |
Mn2+ | pH 7.4, mutant Y140F | Sus scrofa | |
0.000084 | - |
Mn2+ | pH 7.4, mutant Y140E | Sus scrofa | |
0.00011 | - |
Mn2+ | pH 7.4, wild-type enzyme | Sus scrofa | |
0.00042 | - |
NADP+ | pH 7.4, mutant Y140E | Sus scrofa | |
0.00046 | - |
NADP+ | pH 7.4, mutant Y140T | Sus scrofa | |
0.00051 | - |
NADP+ | pH 7.4, mutant Y140K | Sus scrofa | |
0.0006 | - |
isocitrate | pH 7.4, mutant Y140T | Sus scrofa | |
0.0007 | - |
isocitrate | pH 7.4, mutant Y140E | Sus scrofa | |
0.00091 | - |
Mn2+ | pH 7.4, mutant K212Q | Sus scrofa | |
0.0011 | - |
Mn2+ | pH 7.4, mutant K212Y | Sus scrofa | |
0.0023 | - |
NADP+ | pH 7.4, mutant Y140F | Sus scrofa | |
0.0052 | - |
isocitrate | pH 7.4, mutant Y140F | Sus scrofa | |
0.0053 | - |
isocitrate | pH 7.4, mutant Y140K | Sus scrofa | |
0.0073 | - |
NADP+ | pH 7.4, mutant K212Y | Sus scrofa | |
0.0075 | - |
isocitrate | pH 7.4, wild-type enzyme | Sus scrofa | |
0.0092 | - |
isocitrate | pH 7.4, mutant K212Y | Sus scrofa | |
0.0098 | - |
isocitrate | pH 7.4, mutant K212Q | Sus scrofa | |
0.0099 | - |
NADP+ | pH 7.4, wild-type enzyme | Sus scrofa | |
0.014 | - |
NADP+ | pH 7.4, mutant K212Q | Sus scrofa | |
0.018 | - |
NADP+ | pH 7.4, mutant K212R | Sus scrofa | |
0.116 | - |
isocitrate | pH 7.4, mutant K212R | Sus scrofa | |
0.131 | - |
Mn2+ | pH 7.4, mutant Y140K | Sus scrofa | |
0.252 | - |
Mn2+ | pH 7.4, mutant K212R | Sus scrofa |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Sus scrofa | 5739 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | - |
Sus scrofa |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
43000 | - |
2 * 43000, recombinant wild-type and mutant enzymes, SDS-PAGE | Sus scrofa |
92000 | - |
recombinant wild-type and mutant enzymes, native PAGE | Sus scrofa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sus scrofa | P33198 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli, to homogeneity | Sus scrofa |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
isocitrate + NADP+ = 2-oxoglutarate + CO2 + NADPH + H+ | Tyr140 and Lys212 are required for catalytic activity, Tyr140 is the general acid that protonates the substrate after decarboxylation, Lys212 lowers as a positively charged residue the pK of the nearby ionizable group in the enzyme-substrate complex and stabilizes the carbanion formed initially on substrate decarboxylation | Sus scrofa |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
heart | - |
Sus scrofa | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.003 | - |
purified recombinant mutant K212Y, forward reaction | Sus scrofa |
0.065 | - |
purified recombinant mutant K212Q, forward reaction | Sus scrofa |
0.08 | - |
purified recombinant mutant Y140E, forward reaction | Sus scrofa |
0.083 | - |
purified recombinant mutant Y140T, forward reaction | Sus scrofa |
0.102 | - |
purified recombinant mutant Y140F, forward reaction | Sus scrofa |
0.179 | - |
purified recombinant mutant Y140K, forward reaction | Sus scrofa |
3.16 | - |
purified recombinant mutant K212R, forward reaction | Sus scrofa |
35.1 | - |
purified recombinant wild-type enzyme, forward reaction | Sus scrofa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NADP+ | - |
Sus scrofa | 2-oxoglutarate + CO2 + NADPH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 43000, recombinant wild-type and mutant enzymes, SDS-PAGE | Sus scrofa |
Synonyms | Comment | Organism |
---|---|---|
NADP-dependent isocitrate dehydrogenase | - |
Sus scrofa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
forward reaction, assay at | Sus scrofa |
37 | - |
reverse reaction, assay at | Sus scrofa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Sus scrofa |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
pH-profile, recombinant wild-type and mutant enzymes | Sus scrofa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Sus scrofa | |
NADPH | - |
Sus scrofa |