A molecular docking strategy identifies Eosin B as a non-active site inhibitor of protozoal bifunctional thymidylate synthase-dihydrofolate reductase
| Autor: | Chloé E. Atreya, Isabelle Coppens, Antonia Dow, Keith A. Joiner, John J. Irwin, Eric F. Johnson, Karen S. Anderson, Valeska Stempliuk, Stephen M. Beverley, Kristen M. Massimine, Brian K. Shoichet |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Models
Molecular Time Factors Eosine I Bluish Substrate channeling Antiprotozoal Agents Glutamic Acid CHO Cells Biology Arginine Biochemistry Thymidylate synthase Residue (chemistry) chemistry.chemical_compound Inhibitory Concentration 50 Multienzyme Complexes Cricetinae parasitic diseases Dihydrofolate reductase Animals Enzyme Inhibitors Molecular Biology Chromatography High Pressure Liquid Leishmania major chemistry.chemical_classification Binding Sites Dose-Response Relationship Drug Active site Cell Biology Thymidylate Synthase Hydrogen-Ion Concentration Fluoresceins Molecular biology Protein Structure Tertiary Kinetics Tetrahydrofolate Dehydrogenase Enzyme chemistry Eosin B biology.protein Cell culture assays Toxoplasma Protein Binding |
| Zdroj: | The Journal of biological chemistry. 278(16) |
| ISSN: | 0021-9258 |
| Popis: | 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 microm 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 microm inhibitor of Toxoplasma gondii in both biochemical and cell culture assays. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|