Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
to 1.97 A resolution, crystal structure of enzyme bound to pyruvate. Modeling of the 4-hydroxy-2-oxoglutarate-Schiff base intermediate and kinetic analyses of site-directed mutants support the importance of Lys196 as the nucleophile, Tyr168 and Ser77 as components of a proton relay, and Asn78 and Ser198 as unique residues that facilitate substrate binding | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
K196A | complete loss of activity | Homo sapiens |
N78A | 20% of wild-type activity | Homo sapiens |
N78Q | 4% of wild-type activity | Homo sapiens |
N78T | 45% of wild-type activity | Homo sapiens |
S198A | 9% of wild-type activity | Homo sapiens |
S198T | 18% of wild-type activity | Homo sapiens |
S77A | 5% of wild-type activity | Homo sapiens |
S77T | 2% of wild-type activity | Homo sapiens |
S77V | 2% of wild-type activity | Homo sapiens |
Y140F | 122% of wild-type activity | Homo sapiens |
Y168F | complete loss of activity | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.008 | - |
4-hydroxy-2-oxoglutarate | mutant Y140F, pH 8.5, 37°C | Homo sapiens | |
0.011 | - |
4-hydroxy-2-oxoglutarate | wild-type, pH 8.5, 37°C | Homo sapiens | |
0.016 | - |
4-hydroxy-2-oxoglutarate | mutant N78T, pH 8.5, 37°C | Homo sapiens | |
0.044 | - |
4-hydroxy-2-oxoglutarate | mutant S198A, pH 8.5, 37°C | Homo sapiens | |
0.058 | - |
4-hydroxy-2-oxoglutarate | mutant S77T, pH 8.5, 37°C | Homo sapiens | |
0.066 | - |
4-hydroxy-2-oxoglutarate | mutant N78A, pH 8.5, 37°C | Homo sapiens | |
0.077 | - |
4-hydroxy-2-oxoglutarate | mutant S198T, pH 8.5, 37°C | Homo sapiens | |
0.082 | - |
4-hydroxy-2-oxoglutarate | mutant S77V, pH 8.5, 37°C | Homo sapiens | |
0.084 | - |
4-hydroxy-2-oxoglutarate | mutant S77A, pH 8.5, 37°C | Homo sapiens | |
0.281 | - |
4-hydroxy-2-oxoglutarate | mutant N78Q, pH 8.5, 37°C | Homo sapiens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
35249 | - |
and dimer, in equilibrium, 4 * 35249, calculated | Homo sapiens |
35249 | - |
and tetramer, in equilibrium, 2 * 35249, calculated | Homo sapiens |
97200 | - |
sedimentation equilibrium centrifugation | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q86XE5 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-Hydroxy-2-oxoglutarate | - |
Homo sapiens | Pyruvate + glyoxylate | - |
r | |
additional information | enzyme performs a retro-aldol cleavage reaction reminiscent of the trimeric 2-keto-3-deoxy-6-phosphogluconate aldolases | Homo sapiens | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | and tetramer, in equilibrium, 2 * 35249, calculated | Homo sapiens |
tetramer | and dimer, in equilibrium, 4 * 35249, calculated | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.8 | - |
4-hydroxy-2-oxoglutarate | mutant S77T, pH 8.5, 37°C | Homo sapiens | |
1.2 | - |
4-hydroxy-2-oxoglutarate | mutant S77V, pH 8.5, 37°C | Homo sapiens | |
2.4 | - |
4-hydroxy-2-oxoglutarate | mutant S198A, pH 8.5, 37°C | Homo sapiens | |
2.5 | - |
4-hydroxy-2-oxoglutarate | mutant S77A, pH 8.5, 37°C | Homo sapiens | |
4 | - |
4-hydroxy-2-oxoglutarate | mutant N78T, pH 8.5, 37°C | Homo sapiens | |
5.2 | - |
4-hydroxy-2-oxoglutarate | mutant Y140F, pH 8.5, 37°C | Homo sapiens | |
6.1 | - |
4-hydroxy-2-oxoglutarate | wild-type, pH 8.5, 37°C | Homo sapiens | |
6.2 | - |
4-hydroxy-2-oxoglutarate | mutant N78Q, pH 8.5, 37°C | Homo sapiens | |
7.6 | - |
4-hydroxy-2-oxoglutarate | mutant N78A, pH 8.5, 37°C | Homo sapiens | |
7.8 | - |
4-hydroxy-2-oxoglutarate | mutant S198T, pH 8.5, 37°C | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
14 | - |
4-hydroxy-2-oxoglutarate | mutant S77T, pH 8.5, 37°C | Homo sapiens | |
15 | - |
4-hydroxy-2-oxoglutarate | mutant S77V, pH 8.5, 37°C | Homo sapiens | |
22 | - |
4-hydroxy-2-oxoglutarate | mutant N78Q, pH 8.5, 37°C | Homo sapiens | |
30 | - |
4-hydroxy-2-oxoglutarate | mutant S77A, pH 8.5, 37°C | Homo sapiens | |
55 | - |
4-hydroxy-2-oxoglutarate | mutant S198A, pH 8.5, 37°C | Homo sapiens | |
101 | - |
4-hydroxy-2-oxoglutarate | mutant S198T, pH 8.5, 37°C | Homo sapiens | |
115 | - |
4-hydroxy-2-oxoglutarate | mutant N78A, pH 8.5, 37°C | Homo sapiens | |
250 | - |
4-hydroxy-2-oxoglutarate | mutant N78T, pH 8.5, 37°C | Homo sapiens | |
555 | - |
4-hydroxy-2-oxoglutarate | wild-type, pH 8.5, 37°C | Homo sapiens | |
650 | - |
4-hydroxy-2-oxoglutarate | mutant Y140F, pH 8.5, 37°C | Homo sapiens |