Literature summary for 1.5.1.3 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.

Search for more literature for 1.5.1.3

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
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
V115A	mutant resistant to methotrexate, 600-fold increase in Ki value compared with wild-type	Homo sapiens
V115C	mutant resistant to methotrexate, 600-fold increase in Ki value compared with wild-type	Homo sapiens

Inhibitors

Inhibitors	Comment	Organism	Structure
methotrexate	-	Homo sapiens
pemetrexed	-	Homo sapiens

KM Value [mM]

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

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-

Substrates and Products (Substrate)

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

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.65	-	7,8-dihydrofolate	mutant V115C, pH 7.6, 23°C	Homo sapiens
2.8	-	7,8-dihydrofolate	mutant V115A, pH 7.6, 23°C	Homo sapiens
10	-	7,8-dihydrofolate	wild-type, pH 7.6, 23°C	Homo sapiens

Ki Value [mM]

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

IC50 Value

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

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Release 2023.1 (January 2023)