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Literature summary for 1.5.1.3 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)

Cloned (Comment) Organism
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
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
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

Inhibitors Comment Organism Structure
atherospermidine competitive inhibition Leishmania braziliensis
atherospermidine competitive inhibition Leishmania panamensis
atherospermidine competitive inhibition Trypanosoma cruzi
isomoschatoline competitive inhibition Leishmania braziliensis
isomoschatoline competitive inhibition Leishmania panamensis
isomoschatoline competitive inhibition Trypanosoma cruzi
liriodenine competitive inhibition Leishmania braziliensis
liriodenine competitive inhibition Leishmania panamensis
liriodenine competitive inhibition Trypanosoma cruzi
melosmine competitive inhibition Leishmania braziliensis
melosmine competitive inhibition Leishmania panamensis
melosmine competitive inhibition Trypanosoma cruzi
methotrexate competitive inhibition Leishmania braziliensis
methotrexate competitive inhibition Leishmania panamensis
methotrexate competitive inhibition Trypanosoma cruzi
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
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
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
pyrimethamine competitive inhibition Leishmania braziliensis
pyrimethamine competitive inhibition Leishmania panamensis
pyrimethamine competitive inhibition Trypanosoma cruzi
trimethoprim competitive inhibition Leishmania braziliensis
trimethoprim competitive inhibition Leishmania panamensis
trimethoprim competitive inhibition Trypanosoma cruzi

KM Value [mM]

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
7,8-dihydrofolate + NADPH + H+ Trypanosoma cruzi
-
5,6,7,8-tetrahydrofolate + NADP+
-
r
7,8-dihydrofolate + NADPH + H+ Leishmania braziliensis
-
5,6,7,8-tetrahydrofolate + NADP+
-
r
7,8-dihydrofolate + NADPH + H+ Leishmania panamensis
-
5,6,7,8-tetrahydrofolate + NADP+
-
r

Organism

Organism UniProt Comment Textmining
Leishmania braziliensis A4H4P8
-
-
Leishmania panamensis S5M3K7
-
-
Trypanosoma cruzi Q27793
-
-

Purification (Commentary)

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

Storage Stability

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7,8-dihydrofolate + NADPH + H+
-
Trypanosoma cruzi 5,6,7,8-tetrahydrofolate + NADP+
-
r
7,8-dihydrofolate + NADPH + H+
-
Leishmania braziliensis 5,6,7,8-tetrahydrofolate + NADP+
-
r
7,8-dihydrofolate + NADPH + H+
-
Leishmania panamensis 5,6,7,8-tetrahydrofolate + NADP+
-
r

Subunits

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

Synonyms

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

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
assay at Trypanosoma cruzi
6
-
assay at Leishmania braziliensis
6
-
assay at Leishmania panamensis

Cofactor

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

Ki Value [mM]

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

IC50 Value

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

General Information

General Information Comment Organism
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
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
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
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
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
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