title: A FRET-based assay for the discovery of West Nile Virus NS2B-NS3 protease inhibitors.
abstract: The West Nile Virus (WNV) has been a worldwide epidemic since the early 1990s. Currently there are no therapeutic treatments for WNV infections. One particular avenue of treatment is inhibition of the NS2B-NS3 protease, an enzyme that is crucial for WNV replication. In our effort to increase the number of NS2B-NS3 protease inhibitors, we report a novel FRET-based high throughput assay for the discovery of WNV NS2B-NS3 protease inhibitors. For this assay, a FRET-based peptide substrate was synthesized and kinetically characterized with the NS2B-NS3 protease. The new substrate exhibits a K(m) of 3.35 ± 0.31 μM, a k(cat) of 0.0717 ± 0.0016 s(-1) and a k(cat)/K(m) of 21,400 ± 2000 M(-1) s(-1).
authors.affiliation: Department of Chemistry and Biochemistry, California State University, 800 N State College, Fullerton, CA 92834, United States.
authors.forename: Rebecca N
authors.initials: RN
authors.lastname: Adamek
authors.forename: Roxanne V
authors.initials: RV
authors.lastname: Maniquis
authors.forename: Sabaha
authors.initials: S
authors.lastname: Khakoo
authors.forename: Michael D
authors.initials: MD
authors.lastname: Bridges
authors.forename: Nicholas T
authors.initials: NT
authors.lastname: Salzameda
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry* / Q000494:pharmacology*; D055808:Drug Discovery; D057075:Enzyme Assays / Q000379:methods; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006801:Humans; D008958:Models, Molecular; D011480:Protease Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D014901:West Nile Fever / Q000188:drug therapy; D014902:West Nile virus / Q000201:enzymology*
chemical_list: D000998:Antiviral Agents; D011480:Protease Inhibitors
keywords: FRET; Inhibitor assay; NS2B-NS3 protease; West Nile Virus