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Literature summary for 1.5.1.3 extracted from

  • Arrebola, R.; Olmo, A.; Reche, P.; Garvey, E.P.; Santi, D.V.; Ruiz-Perez, L.M.; Gonzalez-Pacanowska, D.
    Isolation and characterization of a mutant dihydrofolate reductase-thymidylate synthase from methotrexate-resistant Leishmania cells (1994), J. Biol. Chem., 269, 10590-10596.
    View publication on PubMed

Application

Application Comment Organism
medicine drug target Leishmania major

Cloned(Commentary)

Cloned (Comment) Organism
cloning of gene encoding mutant R53M enzyme from methotrexate-resistant strain into an expression plasmid, expression in Escherichia coli Leishmania major

Protein Variants

Protein Variants Comment Organism
R53M resistance providing naturally occurring mutant from methotrexate-resistant strain, lower turnover number Leishmania major

Inhibitors

Inhibitors Comment Organism Structure
methotrexate wild-type enzyme, slow binding Leishmania major
additional information binding study of mutant R53M enzyme from methotrexate resistant strain with methotrexate Leishmania major

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0009
-
NADPH wild-type enzyme Leishmania major
0.0012
-
NADPH mutant R53M enzyme Leishmania major
0.0013
-
7,8-dihydrofolate wild-type enzyme Leishmania major
0.0016
-
7,8-dihydrofolate mutant R53M enzyme Leishmania major

Organism

Organism UniProt Comment Textmining
Leishmania major
-
bifunctional enzyme dihydrofolate reductase-thymidylate synthase
-
Leishmania major
-
methotrexate-resistant strain with altered enzyme
-

Source Tissue

Source Tissue Comment Organism Textmining

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
8
-
wild-type enzyme Leishmania major
15
-
mutant R53M enzyme Leishmania major

Substrates and Products (Substrate)

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
7.6
-
7,8-dihydrofolate mutant R53M enzyme Leishmania major
29
-
7,8-dihydrofolate wild-type enzyme Leishmania major

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Leishmania major

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000018
-
methotrexate wild-type enzyme Leishmania major