Cloned (Comment) | Organism |
---|---|
expression of His-tagged mutant V77A/I130M/I138V in Escherichia colii strain M15 | Bacillus anthracis |
recombinant expression of the enzyme | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant mutant V77A/I130M/I138V enzyme in complex with inhibitor 2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine, hanging-drop vapor diffusion in 25% w/v PEG 10,000, 0.1 M MES, pH 6.5, at an equal ratio of protein to crystallization solution. Microseeding is used to obtain isolated crystals in 10% w/v PEG 10000 and 0.1 M MES, pH 6.5, at a protein concentration of 10 mg/ml, improved by 15% ethylene glycol. X-ray diffraction structure determination and analysis at 2.5 A resolution | Bacillus anthracis |
Protein Variants | Comment | Organism |
---|---|---|
V77A/I130M/I138V | site-directed mutagenesis | Bacillus anthracis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2,4-diamino-5-(3-phenylprop-1-ynyl)-6-ethylpyrimidine | - |
Bacillus anthracis | |
2,4-diamino-5-(3-phenylprop-1-ynyl)-6-ethylpyrimidine | - |
Homo sapiens | |
2,4-diamino-5-[3-(2,3-dimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | - |
Bacillus anthracis | |
2,4-diamino-5-[3-(2,3-dimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | - |
Homo sapiens | |
2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | - |
Bacillus anthracis | |
2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | - |
Homo sapiens | |
2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-methylpyrimidine | - |
Bacillus anthracis | |
2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-methylpyrimidine | - |
Homo sapiens | |
2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-npropylpyrimidine | - |
Bacillus anthracis | |
2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-npropylpyrimidine | - |
Homo sapiens | |
2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]pyrimidine | - |
Bacillus anthracis | |
2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]pyrimidine | - |
Homo sapiens | |
2,4-diamino-5-[3-(3,4,5-trimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | - |
Bacillus anthracis | |
2,4-diamino-5-[3-(3,4,5-trimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | - |
Homo sapiens | |
methotrexate | - |
Bacillus anthracis | |
methotrexate | - |
Homo sapiens | |
additional information | inhibitor design based on a trimethoprim-like scaffold, overview | Bacillus anthracis | |
additional information | inhibitor design based on a trimethoprim-like scaffold, overview | Homo sapiens | |
trimethoprim | - |
Bacillus anthracis | |
trimethoprim | - |
Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
7,8-dihydrofolate + NADPH + H+ | Homo sapiens | - |
5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
7,8-dihydrofolate + NADPH + H+ | Bacillus anthracis | - |
5,6,7,8-tetrahydrofolate + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus anthracis | Q81R22 | - |
- |
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Homo sapiens |
recombinant His-tagged mutant V77A/I130M/I138V from Escherichia colii strain M15 by nickel affinity chromatography | Bacillus anthracis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
7,8-dihydrofolate + NADPH + H+ | - |
Homo sapiens | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
7,8-dihydrofolate + NADPH + H+ | - |
Bacillus anthracis | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DHFR | - |
Homo sapiens |
DHFR | - |
Bacillus anthracis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
25 | - |
assay at | Bacillus anthracis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
7 | - |
assay at | Bacillus anthracis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Homo sapiens | |
NADPH | - |
Bacillus anthracis |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.000057 | - |
pH 7.0, 25°C | Homo sapiens | 2,4-diamino-5-[3-(3,4,5-trimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | |
0.00013 | - |
pH 7.0, 25°C | Homo sapiens | 2,4-diamino-5-[3-(2,3-dimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | |
0.00036 | - |
pH 7.0, 25°C | Homo sapiens | 2,4-diamino-5-(3-phenylprop-1-ynyl)-6-ethylpyrimidine | |
0.00089 | - |
pH 7.0, 25°C | Bacillus anthracis | 2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | |
0.00094 | - |
pH 7.0, 25°C | Bacillus anthracis | 2,4-diamino-5-[3-(3,4,5-trimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | |
0.00118 | - |
pH 7.0, 25°C | Homo sapiens | 2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-npropylpyrimidine | |
0.00128 | - |
pH 7.0, 25°C | Homo sapiens | 2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | |
0.0013 | - |
pH 7.0, 25°C | Homo sapiens | 2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-methylpyrimidine | |
0.0013 | - |
pH 7.0, 25°C | Bacillus anthracis | 2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-methylpyrimidine | |
0.0017 | - |
pH 7.0, 25°C | Bacillus anthracis | 2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]pyrimidine | |
0.0032 | - |
pH 7.0, 25°C | Homo sapiens | 2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]pyrimidine | |
0.0032 | - |
pH 7.0, 25°C | Bacillus anthracis | 2,4-diamino-5-(3-phenylprop-1-ynyl)-6-ethylpyrimidine | |
0.0055 | - |
pH 7.0, 25°C | Bacillus anthracis | 2,4-diamino-5-[3-(2,5-dimethoxyphenyl)prop-1-ynyl]-6-npropylpyrimidine | |
0.0092 | - |
pH 7.0, 25°C | Bacillus anthracis | 2,4-diamino-5-[3-(2,3-dimethoxyphenyl)prop-1-ynyl]-6-ethylpyrimidine | |
0.071 | - |
pH 7.0, 25°C | Bacillus anthracis | trimethoprim | |
0.12 | - |
pH 7.0, 25°C | Homo sapiens | trimethoprim |