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Literature summary for 1.5.1.3 extracted from

  • Iwakura, M.; Maki, K.; Takahashi, H.; Takenawa, T.; Yokota, A.; Katayanagi, K.; Kamiyama, T.; Gekko, K.
    Evolutional design of a hyperactive cysteine- and methionine-free mutant of Escherichia coli dihydrofolate reductase (2006), J. Biol. Chem., 281, 13234-13246.
    View publication on PubMed

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant M1A/M16N/M20L/M42Y/M92F/C85A/C152S, increased structural flexibility and an increased size of the N-(p-aminobenzoyl)-L-glutamate binding cleft Escherichia coli

Protein Variants

Protein Variants Comment Organism
C85A/C152S kinetic properties similar to wild-type Escherichia coli
M1A/C85A/C152S kinetic properties similar to wild-type Escherichia coli
M1A/M16F/M20L/M42Y/M92F/C85A/C152S hyperactive mutant, increase in dissociation rate constant of tetrahydrofolate from the enzyme-NADPH-tetrahydrofolate ternary complex Escherichia coli
M1A/M16N/M20L/M42Y/M92F/C85A/C152S hyperactive mutant, increase in dissociation rate constant of tetrahydrofolate from the enzyme-NADPH-tetrahydrofolate ternary complex Escherichia coli
M1A/M16S/M20L/M42Y/M92F/C85A/C152S hyperactive mutant, increase in dissociation rate constant of tetrahydrofolate from the enzyme-NADPH-tetrahydrofolate ternary complex Escherichia coli
M1P/C85A/C152S kinetic properties similar to wild-type Escherichia coli
M1S/C85A/C152S kinetic properties similar to wild-type Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0012
-
7,8-dihydrofolate mutant C85A/C152S, 15°C, pH 7.0 Escherichia coli
0.0012
-
7,8-dihydrofolate mutant M1P/C85A/C152S, 15°C, pH 7.0 Escherichia coli
0.0012
-
7,8-dihydrofolate wild-type, 15°C, pH 7.0 Escherichia coli
0.0013
-
7,8-dihydrofolate mutant M1A/C85A/C152S, 15°C, pH 7.0 Escherichia coli
0.0025
-
7,8-dihydrofolate mutant M1S/C85A/C152S, 15°C, pH 7.0 Escherichia coli
0.003
-
7,8-dihydrofolate mutant M1A/M16F/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0 Escherichia coli
0.0038
-
7,8-dihydrofolate mutant M1A/M16S/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0 Escherichia coli
0.0083
-
7,8-dihydrofolate mutant M1A/M16N/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0 Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P0ABQ4
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7,8-dihydrofolate + NADPH
-
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
4.6
-
7,8-dihydrofolate mutant M1A/C85A/C152S, 15°C, pH 7.0 Escherichia coli
4.8
-
7,8-dihydrofolate mutant M1P/C85A/C152S, 15°C, pH 7.0 Escherichia coli
4.9
-
7,8-dihydrofolate mutant C85A/C152S, 15°C, pH 7.0 Escherichia coli
5.3
-
7,8-dihydrofolate mutant M1S/C85A/C152S, 15°C, pH 7.0 Escherichia coli
5.3
-
7,8-dihydrofolate wild-type, 15°C, pH 7.0 Escherichia coli
16.2
-
7,8-dihydrofolate mutant M1A/M16S/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0 Escherichia coli
25.8
-
7,8-dihydrofolate mutant M1A/M16F/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0 Escherichia coli
40.5
-
7,8-dihydrofolate mutant M1A/M16N/M20L/M42Y/M92F/C85A/C152S, 15°C, pH 7.0 Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Escherichia coli
NADPH
-
Escherichia coli