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Literature summary extracted from

  • Frey, K.M.; Liu, J.; Lombardo, M.N.; Bolstad, D.B.; Wright, D.L.; Anderson, A.C.
    Crystal structures of wild-type and mutant methicillin-resistant Staphylococcus aureus dihydrofolate reductase reveal an alternate conformation of NADPH that may be linked to trimethoprim resistance (2009), J. Mol. Biol., 387, 1298-1308.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.3 overexpression of His-tagged wild-type and mutant enzmyes in Escherichia coli strain BL21(DE3) as soluble proteins Staphylococcus aureus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.5.1.3 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

EC Number Inhibitors Comment Organism Structure
1.5.1.3 3-(2,4-diamino-6-methylpyrimidin-5-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-yn-1-ol
-
 Staphylococcus aureus
1.5.1.3 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-ethylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]-6-propylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-(2,5-dimethoxyphenyl)prop-1-yn-1-yl]pyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-(2-methoxy-2',6'-dimethylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-(2-methoxy-2'-methylbiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-(2-methoxybiphenyl-4-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-(5-methoxy-2',6'-dimethylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-(5-methoxy-2'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-(5-methoxy-4'-methylbiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-(5-methoxybiphenyl-3-yl)but-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 5-[3-methoxy-3-(3,4,5-trimethoxyphenyl)prop-1-yn-1-yl]-6-methylpyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 6-methyl-5-[3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 6-methyl-5-[3-(3,4,5-trimethoxyphenyl)pent-1-yn-1-yl]pyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 6-methyl-5-[3-methyl-3-(3,4,5-trimethoxyphenyl)but-1-yn-1-yl]pyrimidine-2,4-diamine
-
 Staphylococcus aureus
1.5.1.3 additional information structure based inhibitor development of propargyl derivatives of trimethoprim for inhibition of resistant enzyme, overview Staphylococcus aureus
1.5.1.3 trimethoprim inhibition of the wild-type enzyme, but 74fold less of the resistant mutant F98Y Staphylococcus aureus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH + H+ Staphylococcus aureus
-
 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Staphylococcus aureus P0A016
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.3 recombinant His-tagged wild-type and mutant enzmyes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Staphylococcus aureus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH + H+
-
 Staphylococcus aureus 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.5.1.3 DHFR
-
 Staphylococcus aureus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.1.3 25
-
 assay at Staphylococcus aureus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.1.3 7
-
 assay at Staphylococcus aureus

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.3 NADPH binding structure and comformation of wild-type and mutant enzyme Staphylococcus aureus

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.5.1.3 0.000023
-
 pH 7.0, 25°C, wild-type enzyme Staphylococcus aureus trimethoprim
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 0.0017
-
 pH 7.0, 25°C, mutant F98Y Staphylococcus aureus trimethoprim
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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
1.5.1.3 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