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Literature summary extracted from

  • Trimmer, E.E.; Ballou, D.P.; Galloway, L.J.; Scannell, S.A.; Brinker, D.R.; Casas, K.R.
    Aspartate 120 of Escherichia coli methylenetetrahydrofolate reductase: evidence for major role in folate binding and catalysis and a minor role in flavin reactivity (2005), Biochemistry, 44, 6809-6822.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.20 expression of wild-type and mutant enzymes in Escherichia coli strain AB109 and AB1909-(DE3)7D Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
1.5.1.20 D120A site-directed mutagenesis, decreased catalytic efficiency in the folate oxidative half-reaction, loss of negative charge near the flavin, small increasing effects on the NADH reductive half reaction Escherichia coli
1.5.1.20 D120K site-directed mutagenesis, decreased catalytic efficiency in the folate oxidative half-reaction, loss of negative charge near the flavin, small increasing effects on the NADH reductive half reaction Escherichia coli
1.5.1.20 D120N site-directed mutagenesis, decreased catalytic efficiency in the folate oxidative half-reaction, loss of negative charge near the flavin, small increasing effects on the NADH reductive half reaction Escherichia coli
1.5.1.20 D120S site-directed mutagenesis, decreased catalytic efficiency in the folate oxidative half-reaction, loss of negative charge near the flavin, small increasing effects on the NADH reductive half reaction Escherichia coli
1.5.1.20 D120V site-directed mutagenesis, decreased catalytic efficiency in the folate oxidative half-reaction, loss of negative charge near the flavin, small increasing effects on the NADH reductive half reaction Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.20 methylenetetrahydrofolate
-
Escherichia coli
1.5.1.20 NADH
-
Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.20 additional information
-
additional information redox potentials of wild-type and mutant enzymes, kinetic mechanism, and rapid-reaction kinetics for the half-reactions, steady-state kinetics Escherichia coli
1.5.1.20 0.0004
-
methylenetetrahydrofolate recombinant wild-type enzyme, pH 7.2, 4°C Escherichia coli
1.5.1.20 0.0035
-
NADH recombinant wild-type enzyme, pH 7.2, 4°C, the mutant enzymes all show a Km below 0.0035 mM Escherichia coli
1.5.1.20 0.017
-
methylenetetrahydrofolate recombinant mutant D120N, pH 7.2, 4°C Escherichia coli
1.5.1.20 0.043
-
methylenetetrahydrofolate recombinant mutant D120S, pH 7.2, 4°C Escherichia coli
1.5.1.20 0.099
-
methylenetetrahydrofolate recombinant mutant D120A, pH 7.2, 4°C Escherichia coli
1.5.1.20 0.142
-
methylenetetrahydrofolate recombinant mutant D120K, pH 7.2, 4°C Escherichia coli
1.5.1.20 0.187
-
methylenetetrahydrofolate recombinant mutant D120V, pH 7.2, 4°C Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.1.20 5-methyltetrahydrofolate + NAD+ Escherichia coli
-
5,10-methylenetetrahydrofolate + NADH
-
r

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.20 Escherichia coli
-
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
1.5.1.20 5-methyltetrahydrofolate + NAD(P)+ = 5,10-methylenetetrahydrofolate + NAD(P)H + H+ substrate binding and catalyic mechanism involve Asp120, half-reaction mechanisms Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.20 (6R,S)-5,10-methylenetetrahydrofolate + ?
-
Escherichia coli ?
-
?
1.5.1.20 5-methyltetrahydrofolate + NAD+
-
Escherichia coli 5,10-methylenetetrahydrofolate + NADH
-
r

Synonyms

EC Number Synonyms Comment Organism
1.5.1.20 methylenetetrahydrofolate reductase
-
Escherichia coli
1.5.1.20 MTHFR
-
Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.1.20 0.007
-
methylenetetrahydrofolate recombinant mutant D120N, pH 7.2, 4°C Escherichia coli
1.5.1.20 0.011
-
methylenetetrahydrofolate recombinant mutant D120K, pH 7.2, 4°C Escherichia coli
1.5.1.20 0.017
-
methylenetetrahydrofolate recombinant mutant D120V, pH 7.2, 4°C Escherichia coli
1.5.1.20 0.02
-
methylenetetrahydrofolate recombinant mutant D120S, pH 7.2, 4°C Escherichia coli
1.5.1.20 0.022
-
methylenetetrahydrofolate recombinant mutant D120A, pH 7.2, 4°C Escherichia coli
1.5.1.20 2.2
-
methylenetetrahydrofolate recombinant wild-type enzyme, pH 7.2, 4°C Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.1.20 7.2
-
assay at Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.20 FAD redox properties of enzyme-bound FAD are influenced by Asp120, which electrostatically stabilizes putative 5-iminium cation intermediate during catalysis Escherichia coli
1.5.1.20 NAD+
-
Escherichia coli
1.5.1.20 NADH
-
Escherichia coli

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.5.1.20 0.014
-
NADH recombinant wild-type enzyme, pH 7.2, 4°C, the mutant enzymes all show a Km below 0.0035 mM Escherichia coli
1.5.1.20 0.061
-
methylenetetrahydrofolate recombinant wild-type enzyme, pH 7.2, 4°C Escherichia coli