Crystallization (Comment) | Organism |
---|---|
structures of the enzyme complexes of wild-type with NADH and mutant E28Q with methytetrahydrofolate. Residue Gln183 makes key hydrogen-bonding interactions with both NADH and folate in their respective halfreactions | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
Q183A | in the reductive half-reaction, NADH binding affinity and the rate of flavin reduction are not hindered by the mutation. Q183A exhibits a 6-10fold lower rate of folate reduction and binds methylentetrahydrofolate with 7-fold lower affinity | Escherichia coli |
Q183E | in the reductive half-reaction, NADH binding affinity and the rate of flavin reduction are not hindered by the mutation and Gln183 plays a minor in the oxidative half-reaction. The mutant displays catalytic constants within 3fold of the wild-type enzyme | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5,10-methylenetetrahydrofolate | substrate inhibition | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0004 | - |
5,10-methylenetetrahydrofolate | wild-type, pH 7.2, 4°C | Escherichia coli | |
0.0035 | - |
NADH | wild-type, pH 7.2, 4°C | Escherichia coli | |
0.0066 | - |
NADH | mutant Q183E, pH 7.2, 4°C | Escherichia coli | |
0.01 | - |
NADH | mutant Q183A, pH 7.2, 4°C | Escherichia coli | |
0.104 | - |
5,10-methylenetetrahydrofolate | mutant Q183A, pH 7.2, 4°C | Escherichia coli | |
0.108 | - |
5,10-methylenetetrahydrofolate | mutant Q183E, pH 7.2, 4°C | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0AEZ1 | cf. EC 1.5.1.20 | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5,10-methylenetetrahydrofolate + NADH + H+ | - |
Escherichia coli | 5-methyltetrahydrofolate + NAD+ | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.29 | - |
NADH | mutant Q183A, pH 7.2, 4°C | Escherichia coli | |
2.2 | - |
NADH | wild-type, pH 7.2, 4°C | Escherichia coli | |
2.7 | - |
NADH | mutant Q183E, pH 7.2, 4°C | Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.061 | - |
5,10-methylenetetrahydrofolate | wild-type, pH 7.2, 4°C | Escherichia coli | |
0.094 | - |
5,10-methylenetetrahydrofolate | mutant Q183E, pH 7.2, 4°C | Escherichia coli |