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Literature summary extracted from

  • Tahar, R.; de Pecoulas, P.E.; Basco, L.K.; Chiadmi, M.; Mazabraud, A.
    Kinetic properties of dihydrofolate reductase from wild-type and mutant Plasmodium vivax expressed in Escherichia coli (2001), Mol. Biochem. Parasitol., 113, 241-249.
    View publication on PubMed

Application

EC Number Application Comment Organism
1.5.1.3 medicine enzyme is a target of several anti-folate inhibitory drugs to combat bacteria, protozoa and cancer Plasmodium vivax

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.3 DNA-sequence analysis Plasmodium vivax
1.5.1.3 expression in Escherichia coli of wild-type and mutant enzymes Plasmodium vivax

Protein Variants

EC Number Protein Variants Comment Organism
1.5.1.3 S58R/S117N double mutant with naturally occurring point mutations, antifolate drug resistant Plasmodium vivax

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.3 cycloguanil wild-type and mutant Plasmodium vivax
1.5.1.3 methotrexate wild-type and mutant Plasmodium vivax
1.5.1.3 pyrimethamine wild-type and mutant Plasmodium vivax
1.5.1.3 trimethoprim wild-type and mutant Plasmodium vivax

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 0.06
-
 NADPH wild-type enzyme Plasmodium vivax
1.5.1.3 0.07
-
 NADPH mutant enzyme Plasmodium vivax
1.5.1.3 0.097
-
 7,8-dihydrofolate mutant enzyme Plasmodium vivax
1.5.1.3 0.107
-
 7,8-dihydrofolate wild-type enzyme Plasmodium vivax

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH Plasmodium vivax DHFR activity is not essential for virus replication 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Plasmodium vivax
-
 wild-type and mutant enzymes recombinant from Escherichia coli
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.3 recombinant from Escherichia coli Plasmodium vivax

Renatured (Commentary)

EC Number Renatured (Comment) Organism
1.5.1.3 wild-type and mutant, renaturation from inclusion bodies due to expression in E. coli, 200 mM KCl, 20 mM potassium phosphate, pH 7.0, 0.1 mM EDTA, 10 mM dithiothreitol Plasmodium vivax

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.5.1.3 1.7
-
 purified recombinant enzyme Plasmodium vivax

Storage Stability

EC Number Storage Stability Organism
1.5.1.3 -20°C, several months Plasmodium vivax

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH
-
 Plasmodium vivax 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
1.5.1.3 7,8-dihydrofolate + NADPH DHFR activity is not essential for virus replication Plasmodium vivax 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.1.3 25
-
 assay at Plasmodium vivax

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.3 NADPH
-
 Plasmodium vivax

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.5.1.3 0.0000052
-
 methotrexate wild-type enzyme Plasmodium vivax
1.5.1.3 0.0000069
-
 methotrexate mutant enzyme Plasmodium vivax
1.5.1.3 0.000064
-
 pyrimethamine wild-type enzyme Plasmodium vivax
1.5.1.3 0.000067
-
 cycloguanil wild-type enzyme Plasmodium vivax
1.5.1.3 0.000098
-
 trimethoprim wild-type enzyme Plasmodium vivax
1.5.1.3 0.000143
-
 trimethoprim mutant enzyme Plasmodium vivax
1.5.1.3 0.000743
-
 cycloguanil mutant enzyme Plasmodium vivax
1.5.1.3 0.00093
-
 pyrimethamine mutant enzyme Plasmodium vivax