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Literature summary extracted from

  • Raymond, R.K.; Kastanos, E.K.; Appling, D.R.
    Saccharomyces cerevisiae expresses two genes encoding isoenzymes of methylenetetrahydrofolate reductase (1999), Arch. Biochem. Biophys., 372, 300-308.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.20 cloning, disruption and expression of the two genes MET12 and MET13 encoding isoenzymes of MTHFR, MET12 is located on chromosome XVI and encodes a 657 amino acids protein, MET13 is located on chromosome VII, encodes a 599 amino acids protein and is responsible for most of the activity in cell, expression in Escherichia coli BL21 Saccharomyces cerevisiae

Protein Variants

EC Number Protein Variants Comment Organism
1.5.1.20 additional information mutant strain RRY1 with single disruption of MET13 and strain RRY3 with double disruption of both MET12 and MET13, but not strain RRY2 with single disruption of MET12, result in loss of MTHFR activity and methionine auxothrophy, they can be complemented by overexpression of the human enzyme, but not by overexpression of the Escherichia coli metF gene Saccharomyces cerevisiae

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.20 menadione inhibits enzyme activity in yeast extracts Saccharomyces cerevisiae
1.5.1.20 additional information gene contains a putative S-adenosylmethionione binding sequence, that is inhibitory in eukaryotic MTHFR Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.1.20 5,10-methylenetetrahydrofolate + NADPH Saccharomyces cerevisiae enzyme encoded by MET13 produces 5-methyltetrahydrofolate used for methylation of homocysteine for methionine biosynthesis in vivo, NADPH is the likely natural reductant 5-methyltetrahydrofolate + NADP+
-
 ?
1.5.1.20 5,10-methylenetetrahydrofolate + NADPH Saccharomyces cerevisiae DAY4 enzyme encoded by MET13 produces 5-methyltetrahydrofolate used for methylation of homocysteine for methionine biosynthesis in vivo, NADPH is the likely natural reductant 5-methyltetrahydrofolate + NADP+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.20 Saccharomyces cerevisiae
-
 two isoenzymes, encoded by the genes MET12 and MET13
-
1.5.1.20 Saccharomyces cerevisiae DAY4
-
 two isoenzymes, encoded by the genes MET12 and MET13
-
1.5.1.20 Schizosaccharomyces pombe
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.5.1.20 0.000094
-
 wild-type, cell extract Saccharomyces cerevisiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.20 5,10-methylenetetrahydrofolate + NADPH enzyme encoded by MET13 produces 5-methyltetrahydrofolate used for methylation of homocysteine for methionine biosynthesis in vivo, NADPH is the likely natural reductant Saccharomyces cerevisiae 5-methyltetrahydrofolate + NADP+
-
 ?
1.5.1.20 5,10-methylenetetrahydrofolate + NADPH enzyme encoded by MET13 produces 5-methyltetrahydrofolate used for methylation of homocysteine for methionine biosynthesis in vivo, NADPH is the likely natural reductant Saccharomyces cerevisiae DAY4 5-methyltetrahydrofolate + NADP+
-
 ?
1.5.1.20 5,10-methylenetetrahydrofolate + reduced acceptor equilibrium lies far in favor of 5-methyltetrahydrofolate formation Saccharomyces cerevisiae 5-methyltetrahydrofolate + oxidized acceptor
-
 r
1.5.1.20 5,10-methylenetetrahydrofolate + reduced acceptor forward reaction: NADPH as reduced acceptor Saccharomyces cerevisiae 5-methyltetrahydrofolate + oxidized acceptor
-
 r
1.5.1.20 5,10-methylenetetrahydrofolate + reduced acceptor equilibrium lies far in favor of 5-methyltetrahydrofolate formation Saccharomyces cerevisiae DAY4 5-methyltetrahydrofolate + oxidized acceptor
-
 r
1.5.1.20 5,10-methylenetetrahydrofolate + reduced acceptor forward reaction: NADPH as reduced acceptor Saccharomyces cerevisiae DAY4 5-methyltetrahydrofolate + oxidized acceptor
-
 r
1.5.1.20 additional information reverse reaction: menadione and 2,6-dichlorophenolindophenol and NADP+ are not effective as electron acceptor Saccharomyces cerevisiae ?
-
 ?
1.5.1.20 additional information reverse reaction: menadione and 2,6-dichlorophenolindophenol and NADP+ are not effective as electron acceptor Saccharomyces cerevisiae DAY4 ?
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.1.20 37
-
 assay at Saccharomyces cerevisiae

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.20 flavin flavin-dependent enzyme Saccharomyces cerevisiae