Protein Variants | Comment | Organism |
---|---|---|
K352E | mutant with 2fold increased Km-values for folate substrates | Escherichia coli |
K352Q | mutant with 2fold increased Km-values for folate substrates | Escherichia coli |
K352R | no effect on Km-values | Escherichia coli |
K75E | mutant with 2.5fold increased Km-value for 5,10-methylenetetrahydrofolate and 8fold increased Km-value for 5,10-methylenetetrahydropteroyltetraglutamate | Escherichia coli |
K78E | mutant with 1.4fold increased Km-values for folate substrates | Escherichia coli |
K81E | mutant with 3fold increased Km-value for 5,10-methylenetetrahydrofolate and 16fold increased Km-value for 5,10-methylenetetrahydropteroyltetraglutamate | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | values for mutant enzymes | Escherichia coli | |
0.0104 | - |
5,10-methylenetetrahydropteroyltetraglutamate | - |
Escherichia coli | |
0.0677 | - |
5,10-methylenetetrahydrofolate | - |
Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
recombinant enzyme, expressed in Escherichia coli BL21 (DE3)pLysS | - |
Purification (Comment) | Organism |
---|---|
purification of recombinant enzyme, expressed in Escherichia coli BL21 (DE3)pLysS, and of mutants | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | enzyme is a component of the glycine cleavage system which is composed of P-, H-, L- and T-protein, multienzyme complex | Escherichia coli | ? | - |
? | |
S-aminomethyldihydrolipoylprotein + (6S)-tetrahydrofolate | aminomethyl intermediate bound to the lipoate cofactor of H-protein | Escherichia coli | dihydrolipoylprotein + (6R)-5,10-methylenetetrahydrofolate + NH3 | - |
r | |
S-aminomethyldihydrolipoylprotein + (6S)-tetrahydrofolate | T-protein catalyzes the tetrahydrofolate-dependent step of the glycine cleavage reaction | Escherichia coli | dihydrolipoylprotein + (6R)-5,10-methylenetetrahydrofolate + NH3 | - |
r | |
S-aminomethyldihydrolipoylprotein + (6S)-tetrahydrofolate | tetrahydrofolate-dependent enzyme | Escherichia coli | dihydrolipoylprotein + (6R)-5,10-methylenetetrahydrofolate + NH3 | - |
r | |
S-aminomethyldihydrolipoylprotein + (6S)-tetrahydrofolate | folate-binding site: Lys-78, Lys-81 and Lys-352 are involved in binding, Lys-352 may serve as the primary binding site to alpha-carboxyl group of the first glutamate residue nearest the p-aminobenzoic acid ring of 5,10-methylenetetrahydrofolate and 5,10-methylenetetrahydropteroyltetraglutamate, Lys-81 may play a key role to hold the second glutamate residue through binding to its alpha-carboxyl group, 6.5fold higher affinity for 5,10-methylenetetrahydropteroyltetraglutamate than for 5,10-methylenetetrahydrofolate | Escherichia coli | dihydrolipoylprotein + (6R)-5,10-methylenetetrahydrofolate + NH3 | - |
r | |
S-aminomethyldihydrolipoylprotein + tetrahydropteroyltetraglutamate | - |
Escherichia coli | dihydrolipoylprotein + 5,10-methylenetetrahydropteroyltetraglutamate + NH3 | better substrate than 5,10-methylenetetrahydrofolate, 6.5fold higher affinity for 5,10-methylenetetrahydropteroyltetraglutamate than for 5,10-methylenetetrahydrofolate | ? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | values for mutant enzymes | Escherichia coli | |
8.75 | - |
5,10-methylenetetrahydropteroyltetraglutamate | - |
Escherichia coli | |
14.4 | - |
5,10-methylenetetrahydrofolate | - |
Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
tetrahydrofolate | folate-binding site | Escherichia coli | |
tetrahydrofolate | tetrahydrofolate-dependent enzyme | Escherichia coli |