A FRET-based assay for the discovery of West Nile Virus NS2B-NS3 protease inhibitors

作者：Adamek Rebecca N; Maniquis Roxanne V; Khakoo Sabaha; Bridges Michael D; Salzameda Nicholas T^*

来源：Bioorganic & Medicinal Chemistry Letters, 2013, 23(17): 4848-4850.

DOI：10.1016/j.bmcl.2013.06.081

• West Nile Virus
• NS2B-NS3 protease
• Inhibitor assay
• FRET

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 mu M, a k(cat) of 0.0717 +/- 0.0016 s(-1) and a k(cat)/K-m of 21,400 +/- 2000 M-1 s(-1).

• 出版日期2013-9-1