Crystallization (Comment) | Organism |
---|---|
ligand-free mutant F223L and mutant F223L/E28Q in complex with methylenetetrahydrofolate, to 1.65 and 1.7 A resolution, respectively. folate is bound in a catalytically competent conformation, and L223 undergoes a conformational change similar to that observed for F223 in the E28Q-methylenetetrahydrofolate structure | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
F223A | mutation impairs both NADH and methylenetetrahydrofolate binding each 40fold and slows catalysis of both half-reactins less than 2fold | Escherichia coli |
F223L | affinity for methylenetetrahydrofolate is unaffeacted. Mutant catalyzes the oxidative half-reaction 3fold faster than wild-type | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5,10-methylenetetrahydrofolate | - |
Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0005 | - |
5,10-methylenetetrahydrofolate | wild-type, pH 7.2, 25°C | Escherichia coli | |
0.008 | - |
5,10-methylenetetrahydrofolate | mutant F223L, pH 7.2, 25°C | Escherichia coli | |
0.02 | - |
NADH | cosubstrate 5,10-methylenetetrahydrofolate, wild-type, pH 7.2, 25°C | Escherichia coli | |
0.066 | - |
NADH | cosubstrate menadione, wild-type, pH 7.2, 25°C | Escherichia coli | |
0.093 | - |
5,10-methylenetetrahydrofolate | mutant F223A, pH 7.2, 25°C | Escherichia coli | |
0.14 | - |
NADH | cosubstrate 5,10-methylenetetrahydrofolate, mutant F223A, pH 7.2, 25°C | Escherichia coli | |
0.236 | - |
NADH | cosubstrate 5,10-methylenetetrahydrofolate, mutant F223L, pH 7.2, 25°C | Escherichia coli | |
0.47 | - |
NADH | cosubstrate menadione, mutant F223L, pH 7.2, 25°C | Escherichia coli | |
0.585 | - |
NADH | cosubstrate menadione, mutant F223A, pH 7.2, 25°C | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0AEZ1 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5,10-methylenetetrahydrofolate + NADH | - |
Escherichia coli | 5-methyltetrahydrofolate + NAD+ | - |
? | |
NADH + menadione | - |
Escherichia coli | NAD+ + reduced menadione | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
10.4 | - |
NADH | cosubstrate 5,10-methylenetetrahydrofolate, wild-type, pH 7.2, 25°C | Escherichia coli | |
14 | - |
NADH | cosubstrate 5,10-methylenetetrahydrofolate, mutant F223L, pH 7.2, 25°C | Escherichia coli | |
21.9 | - |
NADH | cosubstrate 5,10-methylenetetrahydrofolate, mutant F223A, pH 7.2, 25°C | Escherichia coli | |
22 | - |
NADH | cosubstrate menadione, mutant F223A, pH 7.2, 25°C | Escherichia coli | |
31 | - |
NADH | cosubstrate menadione, mutant F223L, pH 7.2, 25°C | Escherichia coli | |
55 | - |
NADH | cosubstrate menadione, wild-type, pH 7.2, 25°C | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Escherichia coli | |
NADH | - |
Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.16 | - |
5,10-methylenetetrahydrofolate | mutant F223L, pH 7.2, 25°C | Escherichia coli | |
0.32 | - |
5,10-methylenetetrahydrofolate | wild-type, pH 7.2, 25°C | Escherichia coli |