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Literature summary extracted from

  • Feng, J.; Goswami, S.; Howell, E.E.
    R67, the other dihydrofolate reductase: rational design of an alternate active site configuration (2008), Biochemistry, 47, 555-565.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.3 expression in Escherichia coli Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
1.5.1.3 K32M weakens binding of dihydrofolate over 60-fold, increases kcat value by a factor of 5 Escherichia coli
1.5.1.3 additional information construction of a mutant isoform that carries the K32M mutation in gene copies 1 and 3 and the Q67H mutation in gene copies 2 and 4 of the tetrameric enzyme. Mutation Q67H can mostly compensate for mutation K32M. In a multimutant that combines the K32M mutation in gene copies 1 and 3 with the Q67H mutation in all gene copies, dihydrolflate is inhibitory, while NADPH is not. A mutant containing only the Q67H substitution in all four gene copies is severely inhibited by both NADPH and substrate Escherichia coli
1.5.1.3 Q67H about 100-fold tightening in binding to both dihydrolfolate and NADPH, mutation can help rescue the K32 M mutation Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 0.0096
-
 7,8-dihydrofolate mutant isoform that carries the K32M mutation in gene copies 1 and 3 and the Q67H mutation in gene copies 2 and 4, pH 7.0, 30°C Escherichia coli
1.5.1.3 0.011
-
 NADPH mutant carrying the K32M mutation in gene copies 1 and 3 with the Q67H mutation in all gene copies, pH 7.0, 30°C Escherichia coli
1.5.1.3 0.014
-
 7,8-dihydrofolate mutant carrying the K32M mutation in gene copies 1 and 3 with the Q67H mutation in all gene copies, pH 7.0, 30°C Escherichia coli
1.5.1.3 0.029
-
 NADPH mutant isoform that carries the K32M mutation in gene copies 1 and 3 and the Q67H mutation in gene copies 2 and 4, pH 7.0, 30°C Escherichia coli

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Escherichia coli
-
 plasmid-encoded isoform R67
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH + H+
-
 Escherichia coli 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.1.3 0.49
-
 7,8-dihydrofolate mutant carrying the K32M mutation in gene copies 1 and 3 with the Q67H mutation in all gene copies, pH 7.0, 30°C Escherichia coli
1.5.1.3 1.7
-
 7,8-dihydrofolate mutant isoform that carries the K32M mutation in gene copies 1 and 3 and the Q67H mutation in gene copies 2 and 4, pH 7.0, 30°C Escherichia coli