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Literature summary extracted from

  • Osorio, E.; Aguilera, C.; Naranjo, N.; Mari­n, M.; Muskus, C.
    Biochemical characterization of the bifunctional enzyme dihydrofolate reductase-thymidylate synthase from Leishmania (Viannia) and its evaluation as a drug target (2013), Biomedica, 33, 393-401 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.3 recombinant bifunctional enzyme DHFR-TS, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain M15(pREP4) Trypanosoma cruzi
1.5.1.3 recombinant bifunctional enzyme DHFR-TS, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain M15(pREP4) Leishmania braziliensis
1.5.1.3 recombinant bifunctional enzyme DHFR-TS, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain M15(pREP4) Leishmania panamensis
2.1.1.45 recombinant bifunctional enzyme DHFR-TS, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain M15(pREP4) Trypanosoma cruzi
2.1.1.45 recombinant bifunctional enzyme DHFR-TS, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain M15(pREP4) Leishmania braziliensis
2.1.1.45 recombinant bifunctional enzyme DHFR-TS, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain M15(pREP4) Leishmania panamensis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.3 atherospermidine competitive inhibition Leishmania braziliensis
1.5.1.3 atherospermidine competitive inhibition Leishmania panamensis
1.5.1.3 atherospermidine competitive inhibition Trypanosoma cruzi
1.5.1.3 isomoschatoline competitive inhibition Leishmania braziliensis
1.5.1.3 isomoschatoline competitive inhibition Leishmania panamensis
1.5.1.3 isomoschatoline competitive inhibition Trypanosoma cruzi
1.5.1.3 liriodenine competitive inhibition Leishmania braziliensis
1.5.1.3 liriodenine competitive inhibition Leishmania panamensis
1.5.1.3 liriodenine competitive inhibition Trypanosoma cruzi
1.5.1.3 melosmine competitive inhibition Leishmania braziliensis
1.5.1.3 melosmine competitive inhibition Leishmania panamensis
1.5.1.3 melosmine competitive inhibition Trypanosoma cruzi
1.5.1.3 methotrexate competitive inhibition Leishmania braziliensis
1.5.1.3 methotrexate competitive inhibition Leishmania panamensis
1.5.1.3 methotrexate competitive inhibition Trypanosoma cruzi
1.5.1.3 additional information inhibitory effect of antifolate drugs on enzymatic activity, and inhibitory activity of four aporphine alkaloids from Rollinia pittieri and Pseudomalmea boyacana, and some 2,4-diamino-pyrimidine antifolates, overview Leishmania braziliensis
1.5.1.3 additional information inhibitory effect of antifolate drugs on enzymatic activity, and inhibitory activity of four aporphine alkaloids from Rollinia pittieri and Pseudomalmea boyacana, and some 2,4-diamino-pyrimidine antifolates, overview Leishmania panamensis
1.5.1.3 additional information inhibitory effect of antifolate drugs on enzymatic activity, and inhibitory activity of four aporphine alkaloids from Rollinia pittieri and Pseudomalmea boyacana, and some 2,4-diamino-pyrimidine antifolates, overview Trypanosoma cruzi
1.5.1.3 pyrimethamine competitive inhibition Leishmania braziliensis
1.5.1.3 pyrimethamine competitive inhibition Leishmania panamensis
1.5.1.3 pyrimethamine competitive inhibition Trypanosoma cruzi
1.5.1.3 trimethoprim competitive inhibition Leishmania braziliensis
1.5.1.3 trimethoprim competitive inhibition Leishmania panamensis
1.5.1.3 trimethoprim competitive inhibition Trypanosoma cruzi
2.1.1.45 atherospermidine competitive inhibition Leishmania braziliensis
2.1.1.45 atherospermidine competitive inhibition Leishmania panamensis
2.1.1.45 atherospermidine competitive inhibition Trypanosoma cruzi
2.1.1.45 isomoschatoline competitive inhibition Leishmania braziliensis
2.1.1.45 isomoschatoline competitive inhibition Leishmania panamensis
2.1.1.45 isomoschatoline competitive inhibition Trypanosoma cruzi
2.1.1.45 liriodenine competitive inhibition Leishmania braziliensis
2.1.1.45 liriodenine competitive inhibition Leishmania panamensis
2.1.1.45 liriodenine competitive inhibition Trypanosoma cruzi
2.1.1.45 melosmine competitive inhibition Leishmania braziliensis
2.1.1.45 melosmine competitive inhibition Leishmania panamensis
2.1.1.45 melosmine competitive inhibition Trypanosoma cruzi
2.1.1.45 methotrexate competitive inhibition Leishmania braziliensis
2.1.1.45 methotrexate competitive inhibition Leishmania panamensis
2.1.1.45 methotrexate competitive inhibition Trypanosoma cruzi
2.1.1.45 additional information inhibitory effect of antifolate drugs on enzymatic activity, and inhibitory activity of four aporphine alkaloids from Rollinia pittieri and Pseudomalmea boyacana, and some 2,4-diamino-pyrimidine antifolates, overview Leishmania braziliensis
2.1.1.45 additional information inhibitory effect of antifolate drugs on enzymatic activity, and inhibitory activity of four aporphine alkaloids from Rollinia pittieri and Pseudomalmea boyacana, and some 2,4-diamino-pyrimidine antifolates, overview Leishmania panamensis
2.1.1.45 additional information inhibitory effect of antifolate drugs on enzymatic activity, and inhibitory activity of four aporphine alkaloids from Rollinia pittieri and Pseudomalmea boyacana, and some 2,4-diamino-pyrimidine antifolates, overview Trypanosoma cruzi
2.1.1.45 pyrimethamine competitive inhibition Leishmania braziliensis
2.1.1.45 pyrimethamine competitive inhibition Leishmania panamensis
2.1.1.45 pyrimethamine competitive inhibition Trypanosoma cruzi
2.1.1.45 trimethoprim competitive inhibition Leishmania braziliensis
2.1.1.45 trimethoprim competitive inhibition Leishmania panamensis
2.1.1.45 trimethoprim competitive inhibition Trypanosoma cruzi

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 additional information
-
 additional information Michaelis-Menten kinetics Leishmania braziliensis
1.5.1.3 0.0556
-
 7,8-dihydrofolate pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis
2.1.1.45 additional information
-
 additional information Michaelis-Menten kinetics Leishmania braziliensis

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 + H+ Trypanosoma cruzi
-
 5,6,7,8-tetrahydrofolate + NADP+
-
 r
1.5.1.3 7,8-dihydrofolate + NADPH + H+ Leishmania braziliensis
-
 5,6,7,8-tetrahydrofolate + NADP+
-
 r
1.5.1.3 7,8-dihydrofolate + NADPH + H+ Leishmania panamensis
-
 5,6,7,8-tetrahydrofolate + NADP+
-
 r
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP Trypanosoma cruzi
-
 dihydrofolate + dTMP
-
 ?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP Leishmania braziliensis
-
 dihydrofolate + dTMP
-
 ?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP Leishmania panamensis
-
 dihydrofolate + dTMP
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Leishmania braziliensis A4H4P8
-
-
1.5.1.3 Leishmania panamensis S5M3K7
-
-
1.5.1.3 Trypanosoma cruzi Q27793
-
-
2.1.1.45 Leishmania braziliensis A4H4P8
-
-
2.1.1.45 Leishmania braziliensis A4H4P8 subsp. viannia
-
2.1.1.45 Leishmania panamensis S5M3K7
-
-
2.1.1.45 Trypanosoma cruzi Q27793
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.3 recombinant His-tagged enzyme from Escherichia coli strain M15(pREP4) by nickel affinity chromatography Trypanosoma cruzi
1.5.1.3 recombinant His-tagged enzyme from Escherichia coli strain M15(pREP4) by nickel affinity chromatography Leishmania braziliensis
1.5.1.3 recombinant His-tagged enzyme from Escherichia coli strain M15(pREP4) by nickel affinity chromatography Leishmania panamensis
2.1.1.45 recombinant His-tagged enzyme from Escherichia coli strain M15(pREP4) by nickel affinity chromatography Trypanosoma cruzi
2.1.1.45 recombinant His-tagged enzyme from Escherichia coli strain M15(pREP4) by nickel affinity chromatography Leishmania braziliensis
2.1.1.45 recombinant His-tagged enzyme from Escherichia coli strain M15(pREP4) by nickel affinity chromatography Leishmania panamensis

Storage Stability

EC Number Storage Stability Organism
1.5.1.3 -20°C, purified recombinant protein, stable at pH 6.0 over a period of 1 month Trypanosoma cruzi
1.5.1.3 -20°C, purified recombinant protein, stable at pH 6.0 over a period of 1 month Leishmania braziliensis
1.5.1.3 -20°C, purified recombinant protein, stable at pH 6.0 over a period of 1 month Leishmania panamensis
2.1.1.45 -20°C, purified recombinant protein, stable at pH 6.0 over a period of 1 month Trypanosoma cruzi
2.1.1.45 -20°C, purified recombinant protein, stable at pH 6.0 over a period of 1 month Leishmania braziliensis
2.1.1.45 -20°C, purified recombinant protein, stable at pH 6.0 over a period of 1 month Leishmania panamensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH + H+
-
 Trypanosoma cruzi 5,6,7,8-tetrahydrofolate + NADP+
-
 r
1.5.1.3 7,8-dihydrofolate + NADPH + H+
-
 Leishmania braziliensis 5,6,7,8-tetrahydrofolate + NADP+
-
 r
1.5.1.3 7,8-dihydrofolate + NADPH + H+
-
 Leishmania panamensis 5,6,7,8-tetrahydrofolate + NADP+
-
 r
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
 Trypanosoma cruzi dihydrofolate + dTMP
-
 ?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
 Leishmania braziliensis dihydrofolate + dTMP
-
 ?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
 Leishmania panamensis dihydrofolate + dTMP
-
 ?

Subunits

EC Number Subunits Comment Organism
1.5.1.3 ? x * 58000, recombinant bifunctional enzyme DHFR-TS, SDS-PAGE Trypanosoma cruzi
1.5.1.3 ? x * 58000, recombinant bifunctional enzyme DHFR-TS, SDS-PAGE Leishmania braziliensis
1.5.1.3 ? x * 58000, recombinant bifunctional enzyme DHFR-TS, SDS-PAGE Leishmania panamensis
2.1.1.45 ? x * 58000, recombinant bifunctional enzyme DHFR-TS, SDS-PAGE Trypanosoma cruzi
2.1.1.45 ? x * 58000, recombinant bifunctional enzyme DHFR-TS, SDS-PAGE Leishmania braziliensis
2.1.1.45 ? x * 58000, recombinant bifunctional enzyme DHFR-TS, SDS-PAGE Leishmania panamensis

Synonyms

EC Number Synonyms Comment Organism
1.5.1.3 DHFR-TS
-
 Trypanosoma cruzi
1.5.1.3 DHFR-TS
-
 Leishmania braziliensis
1.5.1.3 DHFR-TS
-
 Leishmania panamensis
1.5.1.3 dihydrofolate reductase-thymidylate synthase
-
 Trypanosoma cruzi
1.5.1.3 dihydrofolate reductase-thymidylate synthase
-
 Leishmania braziliensis
1.5.1.3 dihydrofolate reductase-thymidylate synthase
-
 Leishmania panamensis
1.5.1.3 LBRM_06_0830
-
 Leishmania braziliensis
2.1.1.45 DHFR-TS
-
 Trypanosoma cruzi
2.1.1.45 DHFR-TS
-
 Leishmania braziliensis
2.1.1.45 DHFR-TS
-
 Leishmania panamensis
2.1.1.45 dihydrofolate reductase-thymidylate synthase
-
 Trypanosoma cruzi
2.1.1.45 dihydrofolate reductase-thymidylate synthase
-
 Leishmania braziliensis
2.1.1.45 dihydrofolate reductase-thymidylate synthase
-
 Leishmania panamensis
2.1.1.45 LBRM_06_0830
-
 Leishmania braziliensis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.1.3 28
-
 assay at Trypanosoma cruzi
1.5.1.3 28
-
 assay at Leishmania braziliensis
1.5.1.3 28
-
 assay at Leishmania panamensis
2.1.1.45 28
-
 assay at Trypanosoma cruzi
2.1.1.45 28
-
 assay at Leishmania braziliensis
2.1.1.45 28
-
 assay at Leishmania panamensis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.1.3 6
-
 assay at Trypanosoma cruzi
1.5.1.3 6
-
 assay at Leishmania braziliensis
1.5.1.3 6
-
 assay at Leishmania panamensis
2.1.1.45 6
-
 assay at Trypanosoma cruzi
2.1.1.45 6
-
 assay at Leishmania braziliensis
2.1.1.45 6
-
 assay at Leishmania panamensis

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.3 NADP+
-
 Trypanosoma cruzi
1.5.1.3 NADP+
-
 Leishmania braziliensis
1.5.1.3 NADP+
-
 Leishmania panamensis
1.5.1.3 NADPH
-
 Trypanosoma cruzi
1.5.1.3 NADPH
-
 Leishmania braziliensis
1.5.1.3 NADPH
-
 Leishmania panamensis

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.5.1.3 additional information
-
 additional information inhibition kinetics Leishmania braziliensis
1.5.1.3 0.022
-
 methotrexate pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis
1.5.1.3 0.033
-
 trimethoprim pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis
1.5.1.3 0.068
-
 pyrimethamine pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis
1.5.1.3 0.717
-
 isomoschatoline pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis
1.5.1.3 0.748
-
 atherospermidine pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis
1.5.1.3 1.26
-
 melosmine pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis
1.5.1.3 2.615
-
 liriodenine pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis
2.1.1.45 additional information
-
 additional information DHFR inhibition kinetics Leishmania braziliensis

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.5.1.3 0.0015
-
 pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis methotrexate
1.5.1.3 0.0125
-
 pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis pyrimethamine
1.5.1.3 0.152
-
 pH 6.0, 28°C, recombinant enzyme Leishmania braziliensis trimethoprim

General Information

EC Number General Information Comment Organism
1.5.1.3 evolution dihydrofolate reductase (DHFR) and thymidylate synthase (TS) have undergone a fusion event generating a single polypeptide but conserving the two functions in trypanosomatids Trypanosoma cruzi
1.5.1.3 evolution dihydrofolate reductase (DHFR) and thymidylate synthase (TS) have undergone a fusion event generating a single polypeptide but conserving the two functions in trypanosomatids Leishmania braziliensis
1.5.1.3 evolution dihydrofolate reductase (DHFR) and thymidylate synthase (TS) have undergone a fusion event generating a single polypeptide but conserving the two functions in trypanosomatids Leishmania panamensis
1.5.1.3 physiological function dihydrofolate reductase is an essential enzyme in the tetrahydrofolate pathway which catalyzes the NADPH-dependent reduction of 7,8-dihydrofolate (H2F) to the 5,6,7,8-tetrahydrofolate needed to maintain intracellular pools of tetrahydrofolate and its derivatives. These are essential cofactors in the biosynthesis of purines, pyrimidines and several amino acids Trypanosoma cruzi
1.5.1.3 physiological function dihydrofolate reductase is an essential enzyme in the tetrahydrofolate pathway which catalyzes the NADPH-dependent reduction of 7,8-dihydrofolate (H2F) to the 5,6,7,8-tetrahydrofolate needed to maintain intracellular pools of tetrahydrofolate and its derivatives. These are essential cofactors in the biosynthesis of purines, pyrimidines and several amino acids Leishmania braziliensis
1.5.1.3 physiological function dihydrofolate reductase is an essential enzyme in the tetrahydrofolate pathway which catalyzes the NADPH-dependent reduction of 7,8-dihydrofolate (H2F) to the 5,6,7,8-tetrahydrofolate needed to maintain intracellular pools of tetrahydrofolate and its derivatives. These are essential cofactors in the biosynthesis of purines, pyrimidines and several amino acids Leishmania panamensis
2.1.1.45 evolution dihydrofolate reductase (DHFR) and thymidylate synthase (TS) have undergone a fusion event generating a single polypeptide but conserving the two functions in trypanosomatids Trypanosoma cruzi
2.1.1.45 evolution dihydrofolate reductase (DHFR) and thymidylate synthase (TS) have undergone a fusion event generating a single polypeptide but conserving the two functions in trypanosomatids Leishmania braziliensis
2.1.1.45 evolution dihydrofolate reductase (DHFR) and thymidylate synthase (TS) have undergone a fusion event generating a single polypeptide but conserving the two functions in trypanosomatids Leishmania panamensis
2.1.1.45 physiological function dihydrofolate reductase is an essential enzyme in the tetrahydrofolate pathway which catalyzes the NADPH-dependent reduction of 7,8-dihydrofolate (H2F) to the 5,6,7,8-tetrahydrofolate needed to maintain intracellular pools of tetrahydrofolate and its derivatives. These are essential cofactors in the biosynthesis of purines, pyrimidines and several amino acids Trypanosoma cruzi
2.1.1.45 physiological function dihydrofolate reductase is an essential enzyme in the tetrahydrofolate pathway which catalyzes the NADPH-dependent reduction of 7,8-dihydrofolate (H2F) to the 5,6,7,8-tetrahydrofolate needed to maintain intracellular pools of tetrahydrofolate and its derivatives. These are essential cofactors in the biosynthesis of purines, pyrimidines and several amino acids Leishmania braziliensis
2.1.1.45 physiological function dihydrofolate reductase is an essential enzyme in the tetrahydrofolate pathway which catalyzes the NADPH-dependent reduction of 7,8-dihydrofolate (H2F) to the 5,6,7,8-tetrahydrofolate needed to maintain intracellular pools of tetrahydrofolate and its derivatives. These are essential cofactors in the biosynthesis of purines, pyrimidines and several amino acids Leishmania panamensis