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Literature summary extracted from

  • Fossati, E.; Volpato, J.P.; Poulin, L.; Guerrero, V.; Dugas, D.A.; Pelletier, J.N.
    2-tier bacterial and in vitro selection of active and methotrexate-resistant variants of human dihydrofolate reductase (2008), J. Biomol. Screen., 13, 504-514.
    View publication on PubMed

Cloned(Commentary)

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

Protein Variants

EC Number Protein Variants Comment Organism
1.5.1.3 additional information streamlined, 2-tier selection and screening protocol to rapidly identify enzyme variants by saturation mutagenesis of active-site residues 7, 15, 24, 70, and 115, each known or suspected to affect ligand binding, The five libraries are selected for conservation of native-like activity and for resistance against methotrexate Homo sapiens
1.5.1.3 V115A mutant resistant to methotrexate, 600-fold increase in Ki value compared with wild-type Homo sapiens
1.5.1.3 V115C mutant resistant to methotrexate, 600-fold increase in Ki value compared with wild-type Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.3 methotrexate
-
 Homo sapiens
1.5.1.3 pemetrexed
-
 Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 0.000075
-
 7,8-dihydrofolate wild-type, pH 7.6, 23°C Homo sapiens
1.5.1.3 0.007
-
 7,8-dihydrofolate mutant V115C, pH 7.6, 23°C Homo sapiens
1.5.1.3 0.015
-
 7,8-dihydrofolate mutant V115A, pH 7.6, 23°C Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Homo sapiens
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH
-
 Homo sapiens 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 0.65
-
 7,8-dihydrofolate mutant V115C, pH 7.6, 23°C Homo sapiens
1.5.1.3 2.8
-
 7,8-dihydrofolate mutant V115A, pH 7.6, 23°C Homo sapiens
1.5.1.3 10
-
 7,8-dihydrofolate wild-type, pH 7.6, 23°C Homo sapiens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.5.1.3 0.000000031
-
 methotrexate wild-type, pH 7.6, 23°C Homo sapiens
1.5.1.3 0.0000005
-
 pemetrexed wild-type, pH 7.6, 23°C Homo sapiens
1.5.1.3 0.00002
-
 methotrexate mutant V115A, pH 7.6, 23°C Homo sapiens
1.5.1.3 0.00002
-
 methotrexate mutant V115C, pH 7.6, 23°C Homo sapiens
1.5.1.3 0.000131
-
 pemetrexed mutant V115A, pH 7.6, 23°C Homo sapiens
1.5.1.3 0.000283
-
 pemetrexed mutant V115C, pH 7.6, 23°C Homo sapiens

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.5.1.3 0.000041
-
 wild-type, pH 7.6, 23°C Homo sapiens methotrexate
1.5.1.3 0.00015
-
 mutant V115A, pH 7.6, 23°C Homo sapiens methotrexate
1.5.1.3 0.00033
-
 mutant V115C, pH 7.6, 23°C Homo sapiens methotrexate
1.5.1.3 0.00075
-
 wild-type, pH 7.6, 23°C Homo sapiens pemetrexed
1.5.1.3 0.001
-
 mutant V115A, pH 7.6, 23°C Homo sapiens pemetrexed
1.5.1.3 0.0043
-
 mutant V115C, pH 7.6, 23°C Homo sapiens pemetrexed