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Literature summary for 2.1.2.10 extracted from

  • Okamura-Ikeda, K.; Fujiwara, K.; Motokawa, Y.
    Identification of the folate binding sites on the Escherichia coli T-protein of the glycine cleavage system (1999), J. Biol. Chem., 274, 17471-17477.
    View publication on PubMed

Protein Variants

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 [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

Organism UniProt Comment Textmining
Escherichia coli
-
recombinant enzyme, expressed in Escherichia coli BL21 (DE3)pLysS
-

Purification (Commentary)

Purification (Comment) Organism
purification of recombinant enzyme, expressed in Escherichia coli BL21 (DE3)pLysS, and of mutants Escherichia coli

Substrates and Products (Substrate)

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Escherichia coli

Turnover Number [1/s]

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

Cofactor Comment Organism Structure
tetrahydrofolate folate-binding site Escherichia coli
tetrahydrofolate tetrahydrofolate-dependent enzyme Escherichia coli