Application | Comment | Organism |
---|---|---|
drug development | dihydrofolate reductase is a potential drug target for the elimination of Brugia malayi, one of the three causative agents of lymphatic filariasis, a parasitic disease | Brugia malayi |
Cloned (Comment) | Organism |
---|---|
sequence comparisons, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Brugia malayi |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2,4-diaminopyrimidine | poor inhibition | Brugia malayi | |
2,4-diaminoquinazoline | low inhibition | Brugia malayi | |
cycloguanil | low inhibition | Brugia malayi | |
methotrexate | strong inhibition | Brugia malayi | |
additional information | antifolate compounds can serve as inhibitors of BmDHFR | Brugia malayi | |
pyrimethamine | - |
Brugia malayi | |
trimethoprim | - |
Brugia malayi |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten steady-state kinetics | Brugia malayi | |
0.0147 | - |
7,8-dihydrofolate | above, pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
7,8-dihydrofolate + NADPH + H+ | Brugia malayi | - |
5,6,7,8-tetrahydrofolate + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Brugia malayi | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Brugia malayi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
7,8-dihydrofolate + NADPH + H+ | - |
Brugia malayi | 5,6,7,8-tetrahydrofolate + NADP+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 22000, recombinant His6-tagged enzyme, SDS-PAGE, x * 22086, sequence calculation, His-tagged enzyme | Brugia malayi |
Synonyms | Comment | Organism |
---|---|---|
BmDHFR | - |
Brugia malayi |
DHFR | - |
Brugia malayi |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Brugia malayi |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.4 | - |
7,8-dihydrofolate | above, pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Brugia malayi |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Brugia malayi | |
NADPH | the equilibrium dissociation constant KD for NADPH is 25nM | Brugia malayi |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | inhibition kinetics | Brugia malayi | |
0.0000013 | - |
methotrexate | pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | |
0.013 | - |
pyrimethamine | pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | |
0.046 | - |
trimethoprim | pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | |
0.065 | - |
2,4-diaminopyrimidine | pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | |
0.459 | - |
2,4-diaminoquinazoline | pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | |
8 | - |
cycloguanil | above, pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.0000031 | - |
pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | methotrexate | |
0.032 | - |
pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | trimethoprim | |
0.109 | - |
pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | pyrimethamine | |
0.154 | - |
pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | 2,4-diaminoquinazoline | |
0.771 | - |
pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | cycloguanil | |
20 | - |
above, pH 6.0, 25°C recombinant His-tagged enzyme | Brugia malayi | 2,4-diaminopyrimidine |
General Information | Comment | Organism |
---|---|---|
additional information | the predicted specificity determining positions in BmDHFR for ligand binding are Val5, Gly12, Met27, Asn44, Ala45, Lys51, Phe60, Val84, Phe89 Leu95, Leu96, Gly133, Ala137, Val139, Phe140, Phe141 and Glu169 | Brugia malayi |