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Literature summary 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

EC Number Protein Variants Comment Organism
2.1.2.10 K352E mutant with 2fold increased Km-values for folate substrates Escherichia coli
2.1.2.10 K352Q mutant with 2fold increased Km-values for folate substrates Escherichia coli
2.1.2.10 K352R no effect on Km-values Escherichia coli
2.1.2.10 K75E mutant with 2.5fold increased Km-value for 5,10-methylenetetrahydrofolate and 8fold increased Km-value for 5,10-methylenetetrahydropteroyltetraglutamate Escherichia coli
2.1.2.10 K78E mutant with 1.4fold increased Km-values for folate substrates Escherichia coli
2.1.2.10 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.2.10 additional information
-
 additional information values for mutant enzymes Escherichia coli
2.1.2.10 0.0104
-
 5,10-methylenetetrahydropteroyltetraglutamate
-
 Escherichia coli
2.1.2.10 0.0677
-
 5,10-methylenetetrahydrofolate
-
 Escherichia coli

Organism

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

Purification (Commentary)

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.2.10 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 ?
-
 ?
2.1.2.10 S-aminomethyldihydrolipoylprotein + (6S)-tetrahydrofolate aminomethyl intermediate bound to the lipoate cofactor of H-protein Escherichia coli dihydrolipoylprotein + (6R)-5,10-methylenetetrahydrofolate + NH3
-
 r
2.1.2.10 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
2.1.2.10 S-aminomethyldihydrolipoylprotein + (6S)-tetrahydrofolate tetrahydrofolate-dependent enzyme Escherichia coli dihydrolipoylprotein + (6R)-5,10-methylenetetrahydrofolate + NH3
-
 r
2.1.2.10 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
2.1.2.10 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]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.2.10 37
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.2.10 additional information
-
 additional information values for mutant enzymes Escherichia coli
2.1.2.10 8.75
-
 5,10-methylenetetrahydropteroyltetraglutamate
-
 Escherichia coli
2.1.2.10 14.4
-
 5,10-methylenetetrahydrofolate
-
 Escherichia coli

Cofactor

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