A molecular docking strategy identifies eosin B as a non-active site inhibitor of protozoal bifunctional thymidylate synthase-dihydrofolate reductase

被引:20
作者
Atreya, CE
Johnson, EF
Irwin, JJ
Dow, A
Massimine, KM
Coppens, I
Stempliuk, V
Beverley, S
Joiner, KA
Shoichet, BK
Anderson, KS
机构
[1] Yale Univ, Sch Med, Dept Pharmacol, New Haven, CT 06520 USA
[2] Yale Univ, Sch Med, Dept Internal Med, Infect Dis Sect, New Haven, CT 06520 USA
[3] Northwestern Univ, Dept Biol Chem & Mol Pharmacol, Chicago, IL 60611 USA
[4] Washington Univ, Sch Med, Dept Mol Microbiol, St Louis, MO 63110 USA
关键词
D O I
10.1074/jbc.M212690200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Protozoal parasites are unusual in that their thymidylate synthase (TS) and dihydrofolate reductase (DHFR) enzymes exist on a single polypeptide. In an effort to probe the possibility of substrate channeling between the TS and DHFR active sites and to identify inhibitors specific for bifunctional TS-DHFR, we used molecular docking to screen for inhibitors targeting the shallow groove connecting the two active sites. Eosin B is a 100 muM non-active site inhibitor of Leishmania major TS-DHFR identified by molecular docking. Eosin B slows both the TS and DHFR reaction rates. When Arg-283, a key residue to which eosin B is predicted to bind, is mutated to glutamate, however, eosin B only minimally inhibits the TS-DHFR reaction. Additionally, eosin B was found to be a 180 muM inhibitor of Toxoplasma gondii in both biochemical and cell culture assays.
引用
收藏
页码:14092 / 14100
页数:9
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