Crystallization (Comment) | Organism |
---|---|
hanging-drop vapour-diffusion method from 2.0-2.2 M ammonium sulfate, 2% PEG 400, and 0.1 M Na-HEPES at pH 6.0. the NADH-bound form of mutant R109L is obtained by soaking a variant crystal in a solution containing 1.22 mM NADH, 2.8 M ammonium sulfate, 2% polyethylene glycol 400 and 0.1 M Na-Hepes at pH 6.0 | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
K167A | extremely weak inhibition by NADH. Does not form hexamers in response to NADH, unlike the wild-type enzyme | Escherichia coli |
R109L | extremely weak inhibition by NADH. Great structural change. Both regions - residue 260-311 and 316-342 - are much less mobile than in wild-type enzyme | Escherichia coli |
R163L | extremely weak inhibition by NADH. Does not form hexamers in response to NADH, unlike the wild-type enzyme | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
NADH | inhibition of wild-type enzyme. Inhibition is extremenly weak in mutant enzymes Y145A, R163L, and K167A | Escherichia coli | |
oxaloacetate | - |
Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
oxaloacetate | mutant enzyme H110A, in presence of 0.1 M KCl | Escherichia coli | |
0.004 | - |
oxaloacetate | mutant enzyme T204A, in presence of 0.1 M KCl | Escherichia coli | |
0.005 | - |
oxaloacetate | mutant enzyme R163L, in presence of 0.1 M KCl | Escherichia coli | |
0.007 | - |
oxaloacetate | mutant enzyme R109L, in presence of 0.1 M KCl | Escherichia coli | |
0.01 | - |
oxaloacetate | mutant enzyme Q182A, in presence of 0.1 M KCl | Escherichia coli | |
0.017 | - |
oxaloacetate | mutant enzyme N189A, in presence of 0.1 M KCl | Escherichia coli | |
0.018 | - |
oxaloacetate | mutant enzyme T111A, in presence of 0.1 M KCl | Escherichia coli | |
0.026 | - |
oxaloacetate | wild-type enzyme, in presence of 0.1 M KCl | Escherichia coli | |
0.037 | - |
oxaloacetate | mutant enzyme K167A, in presence of 0.1 M KCl | Escherichia coli | |
0.049 | - |
acetyl-CoA | mutant enzyme H110A, in presence of 0.1 M KCl | Escherichia coli | |
0.051 | - |
oxaloacetate | mutant enzyme Y145A, in presence of 0.1 M KCl | Escherichia coli | |
0.056 | - |
acetyl-CoA | mutant enzyme Q182A, in presence of 0.1 M KCl | Escherichia coli | |
0.069 | - |
acetyl-CoA | mutant enzyme N189A, in presence of 0.1 M KCl | Escherichia coli | |
0.069 | - |
acetyl-CoA | mutant enzyme R109L, in presence of 0.1 M KCl | Escherichia coli | |
0.079 | - |
acetyl-CoA | mutant enzyme T111A, in presence of 0.1 M KCl | Escherichia coli | |
0.08 | - |
acetyl-CoA | mutant enzyme R163L, in presence of 0.1 M KCl | Escherichia coli | |
0.12 | - |
acetyl-CoA | wild-type enzyme, in presence of 0.1 M KCl | Escherichia coli | |
0.21 | - |
acetyl-CoA | mutant enzyme T204A, in presence of 0.1 M KCl | Escherichia coli | |
0.22 | - |
acetyl-CoA | mutant enzyme K167A, in presence of 0.1 M KCl | Escherichia coli | |
0.23 | - |
acetyl-CoA | mutant enzyme Y145A, in presence of 0.1 M KCl | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + oxaloacetate + H2O | - |
Escherichia coli | citrate + CoA | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | - |
Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
type II citrate synthase | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3 | 6 | acetyl-CoA | mutant enzyme Y145A, in presence of 0.1 M KCl | Escherichia coli | |
3 | 6 | oxaloacetate | mutant enzyme Y145A, in presence of 0.1 M KCl | Escherichia coli | |
54 | - |
acetyl-CoA | mutant enzyme T111A, in presence of 0.1 M KCl | Escherichia coli | |
54 | - |
oxaloacetate | mutant enzyme T111A, in presence of 0.1 M KCl | Escherichia coli | |
67 | - |
acetyl-CoA | mutant enzyme H110A, in presence of 0.1 M KCl | Escherichia coli | |
67 | - |
oxaloacetate | mutant enzyme H110A, in presence of 0.1 M KCl | Escherichia coli | |
81 | - |
acetyl-CoA | wild-type enzyme, in presence of 0.1 M KCl | Escherichia coli | |
81 | - |
oxaloacetate | wild-type enzyme, in presence of 0.1 M KCl | Escherichia coli | |
84 | - |
acetyl-CoA | mutant enzyme K167A, in presence of 0.1 M KCl | Escherichia coli | |
84 | - |
oxaloacetate | mutant enzyme K167A, in presence of 0.1 M KCl | Escherichia coli | |
95 | - |
acetyl-CoA | mutant enzyme Q182A, in presence of 0.1 M KCl | Escherichia coli | |
95 | - |
oxaloacetate | mutant enzyme Q182A, in presence of 0.1 M KCl | Escherichia coli | |
108 | - |
acetyl-CoA | mutant enzyme R163L, in presence of 0.1 M KCl | Escherichia coli | |
108 | - |
oxaloacetate | mutant enzyme R163L, in presence of 0.1 M KCl | Escherichia coli | |
118 | - |
acetyl-CoA | mutant enzyme T204A, in presence of 0.1 M KCl | Escherichia coli | |
118 | - |
oxaloacetate | mutant enzyme T204A, in presence of 0.1 M KCl | Escherichia coli | |
121 | - |
acetyl-CoA | mutant enzyme R109L, in presence of 0.1 M KCl | Escherichia coli | |
121 | - |
oxaloacetate | mutant enzyme R109L, in presence of 0.1 M KCl | Escherichia coli | |
124 | - |
acetyl-CoA | mutant enzyme N189A, in presence of 0.1 M KCl | Escherichia coli | |
124 | - |
oxaloacetate | mutant enzyme N189A, in presence of 0.1 M KCl | Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0017 | - |
NADH | mutant enzyme R109L | Escherichia coli | |
0.0028 | - |
NADH | wild-type enzyme | Escherichia coli | |
0.003 | - |
oxaloacetate | mutant enzyme H110A, in presence of 0.1 M KCl | Escherichia coli | |
0.004 | - |
oxaloacetate | mutant enzyme T204A, in presence of 0.1 M KCl | Escherichia coli | |
0.005 | - |
oxaloacetate | mutant enzyme R163L, in presence of 0.1 M KCl | Escherichia coli | |
0.007 | - |
oxaloacetate | mutant enzyme R109L, in presence of 0.1 M KCl | Escherichia coli | |
0.01 | - |
oxaloacetate | mutant enzyme Q182A, in presence of 0.1 M KCl | Escherichia coli | |
0.017 | - |
oxaloacetate | mutant enzyme N189A, in presence of 0.1 M KCl | Escherichia coli | |
0.018 | - |
NADH | mutant enzyme Q182A | Escherichia coli | |
0.018 | - |
oxaloacetate | mutant enzyme T111A, in presence of 0.1 M KCl | Escherichia coli | |
0.026 | - |
oxaloacetate | wild-type enzyme, in presence of 0.1 M KCl | Escherichia coli | |
0.037 | - |
oxaloacetate | mutant enzyme K167A, in presence of 0.1 M KCl | Escherichia coli | |
0.051 | - |
oxaloacetate | mutant enzyme Y145A, in presence of 0.1 M KCl | Escherichia coli | |
0.08 | - |
NADH | mutant enzyme T111A | Escherichia coli | |
0.121 | - |
NADH | mutant enzyme H110A | Escherichia coli | |
0.165 | - |
NADH | mutant enzyme T204A | Escherichia coli | |
0.242 | - |
NADH | mutant enzyme N189A | Escherichia coli | |
0.4 | - |
NADH | mutant enzyme R163L | Escherichia coli | |
0.63 | - |
NADH | mutant enzyme K167A | Escherichia coli | |
0.79 | - |
NADH | mutant enzyme Y145A | Escherichia coli |