Increased activity of unlinked Zika virus NS2B/NS3 protease compared to linked Zika virus protease

• Published in: Biochemical and biophysical research communications 2017-10, Vol.492 (4), p.668-673
• Main Authors: Kuiper, Benjamin D., Slater, Kristin, Spellmon, Nicholas, Holcomb, Joshua, Medapureddy, Prasanna, Muzzarelli, Kendall M., Yang, Zhe, Ovadia, Reuben, Amblard, Franck, Kovari, Iulia A., Schinazi, Raymond F., Kovari, Ladislau C.
• Format: Article
• Language: English
• Subjects: Enzyme Activation; Enzyme Stability; Flavivirus; NS2B/NS3 protease; Protease inhibitor; Protein Binding; RNA Helicases - chemistry; Serine Endopeptidases - chemistry; Substrate Specificity; Viral Nonstructural Proteins - chemistry; Zika virus; Zika Virus - enzymology
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2017.03.108

Zika virus (ZIKV) is a flavivirus spread by daytime-active Aedes spp. mosquitoes such as A. aegypti and A. albopictus. Previously thought to be a mild infection, the latest ZIKV outbreak in the Americas is causally associated with more severe symptoms as well as severe birth defects, such as microcephaly. Currently no vaccine or antiviral exists. However, recent progress has demonstrated the viral NS2B/NS3 protease may be a suitable target for the development of small-molecule antiviral agents. To better understand the ZIKV protease, we expressed, purified, and characterized unlinked and linked NS2B/NS3 protease corresponding to an isolate from the recent outbreak in Puerto Rico. Unlinked ZIKV protease is more active and binds substrate with greater affinity than linked ZIKV protease. Therefore, we propose that unlinked ZIKV protease be used when evaluating or designing ZIKV protease inhibitors. Additionally, potent inhibitors of related viral proteases, like West Nile Virus and Dengue virus, may serve as advanced starting points to identify and develop ZIKV protease inhibitors. •Unlinked ZIKV protease is more active than linked ZIKV protease.•G4SG4 linker may sterically hinder substrate binding, reduce protease flexibility.•Unlinked ZIKV protease should be used when screening or designing compounds.•Related flavivirus protease inhibitors may be repurposed to target ZIKV protease.