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Literature summary extracted from

  • Nare, B.; Hardy, L.W.; Beverley, S.M.
    The roles of pteridine reductase 1 and dihydrofolate reductase-thymidylate synthase in pteridine metabolism in the protozoan parasite Leishmania major (1997), J. Biol. Chem., 272, 13883-13891.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.3 expression of dhfr gene in Escherichia coli Leishmania major
1.5.1.33 expression in Escherichia coli Leishmania major

Protein Variants

EC Number Protein Variants Comment Organism
1.5.1.3 additional information knockout mutant dhfr-ts- Leishmania major

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.3 methotrexate
-
 Leishmania major
1.5.1.33 2,4-diaminopyrimidines overview Leishmania major
1.5.1.33 dihydrobiopterin substrate inhibition above 0.01 mM Leishmania major
1.5.1.33 dihydrofolate substrate inhibition above 0.005 mM Leishmania major
1.5.1.33 dihydroneopterin substrate inhibition Leishmania major
1.5.1.33 Dihydrosepiapterin substrate inhibition Leishmania major

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 0.0041
-
 folate bifunctional enzyme DHFR-thymidylate synthase Leishmania major
1.5.1.33 0.0016
-
 folate recombinant enzyme, pH 4.7 Leishmania major
1.5.1.33 0.0019
-
 folate
-
 Leishmania tarentolae
1.5.1.33 0.0019
-
 folate recombinant enzyme, pH 6.0 Leishmania major
1.5.1.33 0.0024
-
 folate native enzyme, pH 6.0 Leishmania major
1.5.1.33 0.0024
-
 folate NADPH, with folate as cosubstrate, native enzyme, pH 6.0 Leishmania major
1.5.1.33 0.0026
-
 folate recombinant enzyme, pH 6.0 Leishmania major
1.5.1.33 0.0034
-
 dihydrofolate recombinant enzyme, pH 6.0 Leishmania major
1.5.1.33 0.0054
-
 dihydrobiopterin recombinant enzyme, pH 7.0 Leishmania major
1.5.1.33 0.0054
-
 dihydrobiopterin dihydrofolate, recombinant enzyme, pH 7.0 Leishmania major
1.5.1.33 0.0056
-
 dihydrobiopterin recombinant enzyme, pH 6.0 Leishmania major
1.5.1.33 0.0061
-
 dihydrofolate recombinant enzyme, pH 4.7 Leishmania major
1.5.1.33 0.0067
-
 dihydrofolate recombinant enzyme, pH 6.0 Leishmania tarentolae
1.5.1.33 0.0076
-
 dihydrobiopterin recombinant enzyme, pH 4.7 Leishmania major
1.5.1.33 0.0085
-
 folate recombinant enzyme, pH 7.0 Leishmania major
1.5.1.33 0.0085
-
 folate dihydrobiopterin, recombinant enzyme, pH 4.7 Leishmania tarentolae
1.5.1.33 0.00935
-
 NADPH with dihydrobiopterin as cosubstrate, recombinant enzyme, pH 4.7 Leishmania tarentolae
1.5.1.33 0.0101
-
 biopterin native enzyme, pH 4.7 Leishmania major
1.5.1.33 0.0109
-
 biopterin recombinant enzyme, pH 4.7 Leishmania tarentolae
1.5.1.33 0.0116
-
 biopterin native enzyme, pH 4.7 Leishmania major
1.5.1.33 0.012
-
 NADPH with dihydrofolate as cosubstrate, recombinant enzyme, pH 6.0 Leishmania tarentolae
1.5.1.33 0.0122
-
 biopterin recombinant enzyme, pH 4.7 Leishmania major
1.5.1.33 0.0122
-
 biopterin NADPH, with folate as cosubstrate, recombinant enzyme, pH 6.0 Leishmania major
1.5.1.33 0.0123
-
 NADPH with biopterin as cosubstrate, recombinant enzyme, pH 4.7 Leishmania tarentolae
1.5.1.33 0.0132
-
 NADPH with biopterin as cosubstrate, recombinant enzyme, pH 4.7 Leishmania major
1.5.1.33 0.0142
-
 NADPH with dihydrofolate as cosubstrate, recombinant enzyme, pH 6.0 Leishmania major
1.5.1.33 0.0145
-
 NADPH with dihydrobiopterin as cosubstrate, recombinant enzyme, pH 4.7 Leishmania major
1.5.1.33 0.0146
-
 NADPH with folate as cosubstrate, recombinant enzyme, pH 6.0 Leishmania tarentolae
1.5.1.33 0.0198
-
 biopterin recombinant enzyme, pH 6.0 Leishmania major
1.5.1.33 0.0399
-
 biopterin recombinant enzyme, pH 7.0 Leishmania major

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.5.1.33 30000
-
 4 * 30000, SDS-PAGE Leishmania tarentolae
1.5.1.33 30000
-
 4 * 30000, SDS-PAGE Leishmania major
1.5.1.33 116000
-
 recombinant enzyme, gel filtration Leishmania major
1.5.1.33 117000
-
 native enzyme, gel filtration Leishmania major

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 Leishmania major
-
 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
1.5.1.3 additional information Leishmania major dihydrofolate reductase deficient mutant needs thymidine for growth, pathway overview ?
-
 ?
1.5.1.3 additional information Leishmania major knockout mutants with either no pteridine reductase or dihydrofolate reductase activity revealed in comparison to wild-type that 90% of the cellular dihydrofolate reduction activity belongs to DHFR and 10% of the cellular dihydrofolate reductase activity belongs to pteridine reductase ?
-
 ?
1.5.1.33 additional information Leishmania tarentolae the enzyme mediates the synthesis of tetrahydropteridines ?
-
 ?
1.5.1.33 additional information Leishmania major the enzyme mediates the synthesis of tetrahydropteridines ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Leishmania major
-
 bifunctional enzyme dihydrofolate reductase-thymidylate synthase
-
1.5.1.33 Leishmania major
-
-
-
1.5.1.33 Leishmania tarentolae
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.3 recombinant enzyme from Escherichia coli Leishmania major
1.5.1.33 native and recombinant enzyme Leishmania major

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 0.0025
-
 bifunctional enzyme DHFR-thymidylate synthase, substrate folate Leishmania major
1.5.1.3 6.4
-
 bifunctional enzyme DHFR-thymidylate synthase, substrate 7,8-dihydrofolate Leishmania major

Storage Stability

EC Number Storage Stability Organism
1.5.1.33 -80°C, 20% glycerol, 20 mM beta-mercaptoethanol, stable Leishmania major

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH
-
 Leishmania major 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
1.5.1.3 7,8-dihydrofolate + NADPH bifunctional enzyme DHFR-thymidylate synthase Leishmania major 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
1.5.1.3 folate + NADPH slow rate Leishmania major 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
1.5.1.3 folate + NADPH bifunctional enzyme DHFR-thymidylate synthase Leishmania major 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
1.5.1.3 additional information no activity with pterins Leishmania major ?
-
 ?
1.5.1.3 additional information dihydrofolate reductase deficient mutant needs thymidine for growth, pathway overview Leishmania major ?
-
 ?
1.5.1.3 additional information knockout mutants with either no pteridine reductase or dihydrofolate reductase activity revealed in comparison to wild-type that 90% of the cellular dihydrofolate reduction activity belongs to DHFR and 10% of the cellular dihydrofolate reductase activity belongs to pteridine reductase Leishmania major ?
-
 ?
1.5.1.33 Biopterin + NADPH
-
 Leishmania tarentolae 5,6,7,8-Tetrahydrobiopterin + NADP+
-
 ?
1.5.1.33 Biopterin + NADPH
-
 Leishmania major 5,6,7,8-Tetrahydrobiopterin + NADP+
-
 ?
1.5.1.33 Dihydrobiopterin + NADPH
-
 Leishmania tarentolae ?
-
 ?
1.5.1.33 Dihydrobiopterin + NADPH
-
 Leishmania major ?
-
 ?
1.5.1.33 Dihydrofolate + NADPH
-
 Leishmania tarentolae 5,6,7,8-Tetrahydrofolate + NADP+
-
 ?
1.5.1.33 Dihydrofolate + NADPH
-
 Leishmania major 5,6,7,8-Tetrahydrofolate + NADP+
-
 ?
1.5.1.33 Dihydroneopterin + NADPH
-
 Leishmania tarentolae ?
-
 ?
1.5.1.33 Dihydroneopterin + NADPH
-
 Leishmania major ?
-
 ?
1.5.1.33 dihydrosepiapterin + NADPH
-
 Leishmania tarentolae ?
-
 ?
1.5.1.33 dihydrosepiapterin + NADPH
-
 Leishmania major ?
-
 ?
1.5.1.33 folate + NADPH
-
 Leishmania tarentolae 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
1.5.1.33 folate + NADPH
-
 Leishmania major 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
1.5.1.33 additional information the enzyme mediates the synthesis of tetrahydropteridines Leishmania tarentolae ?
-
 ?
1.5.1.33 additional information the enzyme mediates the synthesis of tetrahydropteridines Leishmania major ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.5.1.33 tetramer 4 * 30000, SDS-PAGE Leishmania tarentolae
1.5.1.33 tetramer 4 * 30000, SDS-PAGE Leishmania major

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.1.3 5
-
 bifunctional enzyme DHFR-thymidylate synthase, substrate folate, sodium acetate buffer Leishmania major
1.5.1.3 7
-
 bifunctional enzyme DHFR-thymidylate synthase, substrate 7,8-dihydrofolate, sodium phosphate buffer Leishmania major
1.5.1.33 4.7
-
 biopterin as substrate Leishmania major
1.5.1.33 4.7
-
 dihydrobiopterin as substrate Leishmania major
1.5.1.33 5 7 dihydrofolate as substrate Leishmania major
1.5.1.33 6
-
 folate as substrate Leishmania major

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.3 NADPH
-
 Leishmania major
1.5.1.33 NADPH dependent on Leishmania tarentolae
1.5.1.33 NADPH dependent on Leishmania major

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.5.1.3 0.0000001
-
 methotrexate recombinant enzyme Leishmania major