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Literature summary for 1.5.1.3 extracted from

  • Vanichtanankul, J.; Taweechai, S.; Yuvaniyama, J.; Vilaivan, T.; Chitnumsub, P.; Kamchonwongpaisan, S.; Yuthavong, Y.
    Trypanosomal dihydrofolate reductase reveals natural antifolate resistance (2011), ACS Chem. Biol., 6, 905-911.
    View publication on PubMed
Search for more literature for 1.5.1.3

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Trypanosoma brucei

Crystallization (Commentary)

Crystallization (Comment) Organism
in complex with NADPH and inhibitors, hanging drop vapor diffusion method, using Trypanosoma brucei

Protein Variants

Protein Variants Comment Organism
T86N the mutant shows reduced catalytic efficiency compared to the wild type enzyme Trypanosoma brucei
T86S the mutant shows reduced catalytic efficiency compared to the wild type enzyme Trypanosoma brucei

Inhibitors

Inhibitors Comment Organism Structure
cycloguanil
-
 Trypanosoma brucei
pyrimethamine
-
 Trypanosoma brucei
WR99210
-
 Trypanosoma brucei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00385
-
 7,8-dihydrofolate wild type enzyme, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.00443
-
 NADPH wild type enzyme, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.00523
-
 NADPH mutant enzyme T86N, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.00792
-
 NADPH mutant enzyme T86S, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.01625
-
 7,8-dihydrofolate mutant enzyme T86S, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.03051
-
 7,8-dihydrofolate mutant enzyme T86N, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
7,8-dihydrofolate + NADPH + H+ Trypanosoma brucei
-
 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Organism

Organism UniProt Comment Textmining
Trypanosoma brucei
-
-
-

Purification (Commentary)

Purification (Comment) Organism
methotrexate-Sepharose column chromatography, SP-Sepharose column chromatography, and Sephacryl S-200 and gel filtration Trypanosoma brucei

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
DHFR
-
 Trypanosoma brucei

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.11
-
 7,8-dihydrofolate wild type enzyme, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
3.75
-
 7,8-dihydrofolate mutant enzyme T86N, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
4.54
-
 7,8-dihydrofolate mutant enzyme T86S, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei

Cofactor

Cofactor Comment Organism Structure
NADPH
-
 Trypanosoma brucei

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0000009
-
 WR99210 mutant enzyme T86S, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.0000011
-
 WR99210 wild type enzyme, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.0000021
-
 pyrimethamine mutant enzyme T86S, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.0000039
-
 WR99210 mutant enzyme T86N, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.0000205
-
 cycloguanil mutant enzyme T86S, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.000024
-
 pyrimethamine wild type enzyme, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.000179
-
 pyrimethamine mutant enzyme T86N, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.000256
-
 cycloguanil wild type enzyme, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
0.001915
-
 cycloguanil mutant enzyme T86N, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
120
-
 7,8-dihydrofolate mutant enzyme T86N, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
280
-
 7,8-dihydrofolate mutant enzyme T86S, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei
810
-
 7,8-dihydrofolate wild type enzyme, in 50 mM TES, pH 7.0, at 37°C Trypanosoma brucei