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Literature summary for 1.5.1.3 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)

Cloned (Comment) Organism
expression in Escherichia coli Escherichia coli

Protein Variants

Protein Variants Comment Organism
K32M weakens binding of dihydrofolate over 60-fold, increases kcat value by a factor of 5 Escherichia coli
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
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
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
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
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

Localization Comment Organism GeneOntology No. Textmining

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
plasmid-encoded isoform R67
-

Substrates and Products (Substrate)

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
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
1.7
-
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