Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, hanging drop vapour diffusion method, 10 mg/ml protein with reservoir solution containing 24% PEG 400, and 0.1 M citrate, pH 4.8, addition of 0.003 ml drops of 0.1 M MgCl2, 5 mM isocitrate or homoisocitrate, and of a 0.001 ml drop 50 mM CdCl2, equilibration against 0.5 ml reservoir solution, 5 days at 20°C, X-ray diffraction structure determination and analysis at 1.85 A resolution, molecular replacement | Thermus thermophilus |
Protein Variants | Comment | Organism |
---|---|---|
V135M | site-directed mutagenesis, tetramer-to-dimer structural transition enhances the activity with isocitrate 1.6fold | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.161 | - |
isocitrate | pH 7.5, 60°C, recombinant mutant V135M | Thermus thermophilus | |
0.521 | - |
isocitrate | pH 7.5, 60°C, recombinant wild-type enzyme | Thermus thermophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Thermus thermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | - |
- |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
homoisocitrate + NAD+ | - |
Thermus thermophilus | ? | - |
? | |
isocitrate + NAD+ | 20fold preferred to homoisocitrate | Thermus thermophilus | ? | - |
? | |
additional information | substrate binding site structure, the substrate specificity is determined by residue Arg85, no activity with 3-isopropylmalate | Thermus thermophilus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | subunit interface, crystal structure analysis | Thermus thermophilus |
Synonyms | Comment | Organism |
---|---|---|
HICDH | - |
Thermus thermophilus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
assay at | Thermus thermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
211 | - |
isocitrate | pH 7.5, 60°C, recombinant mutant V135M | Thermus thermophilus | |
438 | - |
isocitrate | pH 7.5, 60°C, recombinant wild-type enzyme | Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Thermus thermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Thermus thermophilus |