Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
additional information | streamlined, 2-tier selection and screening protocol to rapidly identify enzyme variants by saturation mutagenesis of active-site residues 7, 15, 24, 70, and 115, each known or suspected to affect ligand binding, The five libraries are selected for conservation of native-like activity and for resistance against methotrexate | Homo sapiens |
V115A | mutant resistant to methotrexate, 600-fold increase in Ki value compared with wild-type | Homo sapiens |
V115C | mutant resistant to methotrexate, 600-fold increase in Ki value compared with wild-type | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
methotrexate | - |
Homo sapiens | |
pemetrexed | - |
Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000075 | - |
7,8-dihydrofolate | wild-type, pH 7.6, 23°C | Homo sapiens | |
0.007 | - |
7,8-dihydrofolate | mutant V115C, pH 7.6, 23°C | Homo sapiens | |
0.015 | - |
7,8-dihydrofolate | mutant V115A, pH 7.6, 23°C | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
7,8-dihydrofolate + NADPH | - |
Homo sapiens | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.65 | - |
7,8-dihydrofolate | mutant V115C, pH 7.6, 23°C | Homo sapiens | |
2.8 | - |
7,8-dihydrofolate | mutant V115A, pH 7.6, 23°C | Homo sapiens | |
10 | - |
7,8-dihydrofolate | wild-type, pH 7.6, 23°C | Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000000031 | - |
methotrexate | wild-type, pH 7.6, 23°C | Homo sapiens | |
0.0000005 | - |
pemetrexed | wild-type, pH 7.6, 23°C | Homo sapiens | |
0.00002 | - |
methotrexate | mutant V115A, pH 7.6, 23°C | Homo sapiens | |
0.00002 | - |
methotrexate | mutant V115C, pH 7.6, 23°C | Homo sapiens | |
0.000131 | - |
pemetrexed | mutant V115A, pH 7.6, 23°C | Homo sapiens | |
0.000283 | - |
pemetrexed | mutant V115C, pH 7.6, 23°C | Homo sapiens |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.000041 | - |
wild-type, pH 7.6, 23°C | Homo sapiens | methotrexate | |
0.00015 | - |
mutant V115A, pH 7.6, 23°C | Homo sapiens | methotrexate | |
0.00033 | - |
mutant V115C, pH 7.6, 23°C | Homo sapiens | methotrexate | |
0.00075 | - |
wild-type, pH 7.6, 23°C | Homo sapiens | pemetrexed | |
0.001 | - |
mutant V115A, pH 7.6, 23°C | Homo sapiens | pemetrexed | |
0.0043 | - |
mutant V115C, pH 7.6, 23°C | Homo sapiens | pemetrexed |