Literature summary for 1.5.1.3 extracted from

Bourne, C.R.; Bunce, R.A.; Bourne, P.C.; Berlin, K.D.; Barrow, E.W.; Barrow, W.W.
Crystal structure of Bacillus anthracis dihydrofolate reductase with the dihydrophthalazine-based trimethoprim derivative RAB1 provides a structural explanation of potency and selectivity (2009), Antimicrob. Agents Chemother., 53, 3065-3073.

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression as His-tagged enzyme	Bacillus anthracis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme in complex with inhibitors RAB1 and trimethoprim, DHFR-RAB1 cocrystals grow within 1 to 2 days at 22°C in a sitting-drop format from a 30 mg/ml protein solution mixed 1:1 with a well solution containing 12% PEG 3350, 0.2 M CaCl2, 0.1 M MES, pH 5.8, and 3% glycerol. DHFR-TMP cocrystals grow within 10 days at 22°C in a sitting-drop format from a 42.5 mg/ml protein solution mixed 1:1 with a well solution containing 13% PEG 3350, 0.2 M CaCl2, 0.1 M MES, pH 5.45, and 1% ethanol, X-ray diffraction structure determination and analysis at 2.3 A resolution	Bacillus anthracis

Crystallization (Comment)

Organism

purified recombinant His-tagged enzyme in complex with inhibitors RAB1 and trimethoprim, DHFR-RAB1 cocrystals grow within 1 to 2 days at 22°C in a sitting-drop format from a 30 mg/ml protein solution mixed 1:1 with a well solution containing 12% PEG 3350, 0.2 M CaCl2, 0.1 M MES, pH 5.8, and 3% glycerol. DHFR-TMP cocrystals grow within 10 days at 22°C in a sitting-drop format from a 42.5 mg/ml protein solution mixed 1:1 with a well solution containing 13% PEG 3350, 0.2 M CaCl2, 0.1 M MES, pH 5.45, and 1% ethanol, X-ray diffraction structure determination and analysis at 2.3 A resolution

Bacillus anthracis

Inhibitors

Inhibitors	Comment	Organism
methotrexate	binding structure, overview	Bacillus anthracis
RAB1	i.e. (S,E)-3-(5-((2,4-diaminopyrimidin-5-yl)methyl)-2,3-dimethoxyphenyl)-1-(1-propylphthalazin-2(1H)-yl)prop-2-en-1-one or BAL17662, a dihydrophthalazine-based trimethoprim derivative, synthesis, overview. Binding of RAB1 causes a conformational change of the side chain of Arg58 and Met37 to accommodate the dihydrophthalazine moiety. The dihydrophthalazine group provides a large hydrophobic anchor that embeds within the DHFR active site and accounts for its selective inhibitory activity against Bacillus anthracis, binding structure, overview	Bacillus anthracis
trimethoprim	binding structure, overview	Bacillus anthracis

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Bacillus anthracis	Q81R22	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme by metal affinity chromatography	Bacillus anthracis

Substrates and Products (Substrate)

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

Synonyms

Synonyms	Comment	Organism
DHFR	-	Bacillus anthracis

Cofactor

Cofactor	Comment	Organism	Structure
NADPH	-	Bacillus anthracis

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
0.000015	-	-	Bacillus anthracis	methotrexate
0.000054	-	-	Bacillus anthracis	RAB1
77	-	-	Bacillus anthracis	trimethoprim