Cloned (Comment) | Organism |
---|---|
overexpression of His-tagged wild-type and mutant enzmyes in Escherichia coli strain BL21(DE3) as soluble proteins | Staphylococcus aureus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type and mutant enzymes bound to cofactor NADPH and inhibitors 5, 8, 10, and 15, hanging-drop vaporization method, 12 mg/ml protein is incubated with 1 mM inhibitor and 2 mM NADPH on ice for 2 h, mixing of equal volumes of the protein:ligand:NADPH solution with an optimized crystallization solution consisting of 15% PEG 10000, 150 mM sodium acetate, 100 mM MES, pH 6.5, and 5% butyrlactone, 5-7 days X-ray diffraction structure determination and analysis at 1.69-2.35 A resolutions, overview | Staphylococcus aureus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
3-(2,4-diamino-6-methylpyrimidin-5-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-yn-1-ol | - |
Staphylococcus aureus | |
5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-ethylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-propylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]pyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-(2-methoxy-2',6'-dimethylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-(2-methoxy-2'-methylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-(2-methoxybiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-(5-methoxy-2',6'-dimethylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-(5-methoxy-2'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-(5-methoxy-4'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-(5-methoxybiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
5-[3-methoxy-3-(3,4,5-trimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
6-methyl-5-[3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
6-methyl-5-[3-(3,4,5-trimethoxyphenyl)pent-1-yn-1-yl]pyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
6-methyl-5-[3-methyl-3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine | - |
Staphylococcus aureus | |
additional information | structure based inhibitor development of propargyl derivatives of trimethoprim for inhibition of resistant enzyme, overview | Staphylococcus aureus | |
trimethoprim | inhibition of the wild-type enzyme, but 74fold less of the resistant mutant F98Y | Staphylococcus aureus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
7,8-dihydrofolate + NADPH + H+ | Staphylococcus aureus | - |
5,6,7,8-tetrahydrofolate + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Staphylococcus aureus | P0A016 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzmyes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Staphylococcus aureus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
7,8-dihydrofolate + NADPH + H+ | - |
Staphylococcus aureus | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DHFR | - |
Staphylococcus aureus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Staphylococcus aureus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Staphylococcus aureus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | binding structure and comformation of wild-type and mutant enzyme | Staphylococcus aureus |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.000023 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | trimethoprim | |
0.000042 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(5-methoxybiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.000048 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-propylpyrimidine-2,4-diamine | |
0.000061 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(5-methoxybiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.000064 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 6-methyl-5-[3-methyl-3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine | |
0.000068 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-ethylpyrimidine-2,4-diamine | |
0.000073 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(5-methoxy-2'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.000073 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(5-methoxy-2',6'-dimethylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00012 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 6-methyl-5-[3-(3,4,5-trimethoxyphenyl)pent-1-yn-1-yl]pyrimidine-2,4-diamine | |
0.00017 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(5-methoxybiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00017 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 6-methyl-5-[3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine | |
0.00017 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(2-methoxybiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00018 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(5-methoxy-2',6'-dimethylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00019 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(5-methoxybiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00019 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(5-methoxy-2'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00027 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00027 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]pyrimidine-2,4-diamine | |
0.00041 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(5-methoxy-4'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00041 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(2-methoxy-2',6'-dimethylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.0005 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-(2-methoxy-2'-methylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00055 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-propylpyrimidine-2,4-diamine | |
0.00067 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-ethylpyrimidine-2,4-diamine | |
0.00073 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 5-[3-methoxy-3-(3,4,5-trimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00075 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 6-methyl-5-[3-methyl-3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine | |
0.00077 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(2-methoxybiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00096 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(5-methoxy-4'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.0011 | - |
pH 7.0, 25°C, wild-type enzyme | Staphylococcus aureus | 3-(2,4-diamino-6-methylpyrimidin-5-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-yn-1-ol | |
0.0012 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 6-methyl-5-[3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine | |
0.0013 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.00142 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(2-methoxy-2'-methylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.0017 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | trimethoprim | |
0.0039 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 6-methyl-5-[3-(3,4,5-trimethoxyphenyl)pent-1-yn-1-yl]pyrimidine-2,4-diamine | |
0.0041 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 3-(2,4-diamino-6-methylpyrimidin-5-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-yn-1-ol | |
0.00613 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(2-methoxy-2',6'-dimethylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine | |
0.0069 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]pyrimidine-2,4-diamine | |
0.012 | - |
pH 7.0, 25°C, mutant F98Y | Staphylococcus aureus | 5-[3-methoxy-3-(3,4,5-trimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine |