Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary 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.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.5.1.3 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

EC Number Inhibitors Comment Organism Structure
1.5.1.3 methotrexate binding structure, overview Bacillus anthracis
1.5.1.3 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
1.5.1.3 trimethoprim binding structure, overview Bacillus anthracis

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+ Bacillus anthracis
-
 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Bacillus anthracis Q81R22
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

EC Number Synonyms Comment Organism
1.5.1.3 DHFR
-
 Bacillus anthracis

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.3 NADPH
-
 Bacillus anthracis

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.5.1.3 0.000015
-
-
 Bacillus anthracis methotrexate
1.5.1.3 0.000054
-
-
 Bacillus anthracis RAB1
1.5.1.3 77
-
-
 Bacillus anthracis trimethoprim