Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain TX61 | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, using 2.0 M ammonium sulfate, 0.1 M Tris, pH 8.5, and 20 mM beta-mercaptoethanol | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
A191K | the mutant enzyme displays a kcat value of 6fold lower than wild type | Homo sapiens |
L198P | the mutant enzyme displays a kcat value of 2fold lower than wild type | Homo sapiens |
M190E | the mutant enzyme displays a kcat value of 5.9fold lower than wild type | Homo sapiens |
M190K | the mutant has the value of kcat/Km smaller by a factor of about 7500 than the wild type, the crystal structure of this mutant is similar to that of the wild type with loop 181-197 in the inactive conformation, however, the direct vicinity of the mutation, residues 188-194 of this loop, assumes a different conformation with the positions of Calpha shifted up to 7.2 A, the mutant protein is trapped in an inactive state that does not equilibrate easily with the active conformer | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00085 | - |
dUMP | mutant enzyme A191K, at 37°C | Homo sapiens | |
0.00096 | - |
dUMP | mutant enzyme M190E, at 37°C | Homo sapiens | |
0.00259 | - |
dUMP | wild type enzyme, at 37°C | Homo sapiens | |
0.01563 | - |
dUMP | mutant enzyme L198P, at 37°C | Homo sapiens | |
0.179 | - |
dUMP | mutant enzyme M190K, at 37°C | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P04818 | - |
- |
Purification (Comment) | Organism |
---|---|
Blue-Sepharose column chromatography and Q-Sepharose column chromatography | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5,10-methylenetetrahydrofolate + dUMP | - |
Homo sapiens | dihydrofolate + dTMP | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.03 | - |
dUMP | mutant enzyme M190K, at 37°C | Homo sapiens | |
0.52 | - |
dUMP | mutant enzyme A191K, at 37°C | Homo sapiens | |
0.53 | - |
dUMP | mutant enzyme M190E, at 37°C | Homo sapiens | |
1.64 | - |
dUMP | mutant enzyme L198P, at 37°C | Homo sapiens | |
3.11 | - |
dUMP | wild type enzyme, at 37°C | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
N5,N10-methylenetetrahydrofolate | - |
Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00000016 | - |
dUMP | mutant enzyme M190K, at 37°C | Homo sapiens | |
0.0001 | - |
dUMP | mutant enzyme L198P, at 37°C | Homo sapiens | |
0.00055 | - |
dUMP | mutant enzyme M190E, at 37°C | Homo sapiens | |
0.00061 | - |
dUMP | mutant enzyme A191K, at 37°C | Homo sapiens | |
0.0012 | - |
dUMP | wild type enzyme, at 37°C | Homo sapiens |