Activating Compound | Comment | Organism | Structure |
---|---|---|---|
citrate | 106.29% activity at 1 mM | Phaeodactylum tricornutum |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosetta (DE3) cells | Phaeodactylum tricornutum |
Crystallization (Comment) | Organism |
---|---|
sitting drop vapor diffusion method, using 200 mM NaCl, 100 mM sodium HEPES (pH 7.5), and 25% (w/v) PEG 4000 | Phaeodactylum tricornutum |
Protein Variants | Comment | Organism |
---|---|---|
D459A/N461A/D463A/F465A | the mutant maintains comparable specific activity to that of wild type enzyme | Phaeodactylum tricornutum |
IDH1-EF | the mutant without the EF-hand domain (deleted Gln424-Val495 in a total of 72 residues) completely lost its catalytic activity | Phaeodactylum tricornutum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ADP | 86.63% residual activity at 1 mM | Phaeodactylum tricornutum | |
AMP | 97.88% residual activity at 1 mM | Phaeodactylum tricornutum | |
ATP | 88.7% residual activity at 1 mM | Phaeodactylum tricornutum | |
Ca2+ | 3.87% residual activity at 2 mM | Phaeodactylum tricornutum | |
Co2+ | 5.96% residual activity at 2 mM | Phaeodactylum tricornutum | |
Cu2+ | 5.76% residual activity at 2 mM | Phaeodactylum tricornutum | |
K+ | 4.12% residual activity at 2 mM | Phaeodactylum tricornutum | |
Li+ | 5.15% residual activity at 2 mM | Phaeodactylum tricornutum | |
additional information | not inhibited by 2-oxoglutarate | Phaeodactylum tricornutum | |
Na+ | 4.66% residual activity at 2 mM | Phaeodactylum tricornutum | |
NADH | competitive inhibitor | Phaeodactylum tricornutum | |
Ni2+ | 1.6% residual activity at 2 mM | Phaeodactylum tricornutum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.903 | - |
NAD+ | in the presence of Mg2+, at pH 8.0 and 25°C | Phaeodactylum tricornutum | |
1.1325 | - |
NAD+ | in the presence of Mn2+, at pH 8.0 and 25°C | Phaeodactylum tricornutum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | 2 mM used in assay conditions | Phaeodactylum tricornutum | |
Mn2+ | 2 mM used in assay conditions | Phaeodactylum tricornutum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
110000 | - |
gel filtration | Phaeodactylum tricornutum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NAD+ | Phaeodactylum tricornutum | - |
2-oxoglutarate + CO2 + NADH + H+ | - |
? | |
isocitrate + NAD+ | Phaeodactylum tricornutum FACHB-2174 | - |
2-oxoglutarate + CO2 + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Phaeodactylum tricornutum | - |
- |
- |
Phaeodactylum tricornutum FACHB-2174 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Co2+ affinity column chromatography | Phaeodactylum tricornutum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
78 | - |
mutant enzyme D459A/N461A/D463A/F465A, in the presence of Mn2+, at pH 8.0 and 25°C | Phaeodactylum tricornutum |
87.1 | - |
wild type enzyme, in the presence of Mn2+, at pH 8.0 and 25°C | Phaeodactylum tricornutum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NAD+ | - |
Phaeodactylum tricornutum | 2-oxoglutarate + CO2 + NADH + H+ | - |
? | |
isocitrate + NAD+ | - |
Phaeodactylum tricornutum FACHB-2174 | 2-oxoglutarate + CO2 + NADH + H+ | - |
? | |
additional information | the enzyme has no detectable activity with NADP+ | Phaeodactylum tricornutum | ? | - |
- |
|
additional information | the enzyme has no detectable activity with NADP+ | Phaeodactylum tricornutum FACHB-2174 | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 55000, His6-tagged enzyme, SDS-PAGE | Phaeodactylum tricornutum |
homodimer | 2 * 55100, calculated from amino acid sequence | Phaeodactylum tricornutum |
Synonyms | Comment | Organism |
---|---|---|
IDH1 | - |
Phaeodactylum tricornutum |
NAD+-dependent isocitrate dehydrogenase | - |
Phaeodactylum tricornutum |
NAD-IDH | - |
Phaeodactylum tricornutum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
with Mn2+ | Phaeodactylum tricornutum |
35 | - |
with Mg2+ | Phaeodactylum tricornutum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
28 | 35 | the enzyme is stable below 28°C, but rapidly loses activity above 30°C, and 60% activity remains after a 20 min incubation at 35°C | Phaeodactylum tricornutum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
79 | - |
NAD+ | in the presence of Mg2+, at pH 8.0 and 25°C | Phaeodactylum tricornutum | |
180.5 | - |
NAD+ | in the presence of Mn2+, at pH 8.0 and 25°C | Phaeodactylum tricornutum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
with Mn2+ | Phaeodactylum tricornutum |
8.8 | - |
with Mg2+ | Phaeodactylum tricornutum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Phaeodactylum tricornutum |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.45 | - |
NADH | at pH 8.0 and 25°C | Phaeodactylum tricornutum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
90 | - |
NAD+ | in the presence of Mg2+, at pH 8.0 and 25°C | Phaeodactylum tricornutum | |
160 | - |
NAD+ | in the presence of Mn2+, at pH 8.0 and 25°C | Phaeodactylum tricornutum |