Expression, purification and characterization of a GII.4 Norovirus protease from Minerva virus

• Type of Material: Article
• Authors: Benjamin D. Kuiper, Wayne State University School of Medicine; Kendall M. Muzzarelli, Wayne State University School of Medicine; Bradley J. Keusch, Wayne State University School of Medicine; Joshua Holcomb, Wayne State University School of Medicine; Franck Amblard, Emory University; Peng Liu, Emory University; Shaoman Zhou, Emory University; Iulia A. Kovari, Wayne State University School of Medicine; Zhe Yang, Wayne State University School of Medicine; Raymond Schinazi, Emory University; Ladislau C. Kovari, Wayne State University School of Medicine
• Language: English
• Date: 2018-01-01
• Publisher: Bentham Science
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: 2018
• Final Published Version (URL): http://dx.doi.org/10.2174/1871526518666180521091158
• Title of Journal or Parent Work: Infectious Disorders - Drug Targets
• Volume: 18
• Issue: 3
• Start Page: 224
• End Page: 232
• Grant/Funding Information: RFS is supported in part by NIH grant 1R21 AI-129607 and by the Center for AIDS Research grant 2P30AI050409.
• Keywords: Sequence Analysis, Protein; Enzyme Inhibitors; Norovirus; Escherichia coli; Humans; Polyproteins; Viral Proteins; Potent inhibitor; Polyprotein; Chromatography; Sequence Alignment; Genome, Viral; Drug Design; Purification.; Noroviruses; MVpro inhibitors; Peptide Hydrolases; Benzimidazoles
• Research Categories: Biology, Cell; Biology, Virology; Biology, Genetics

Abstract

Background: Noroviruses are the leading cause of acute gastroenteritis worldwide. Norovirus proteases, which are responsible for cleavage of the viral polyprotein, have become an attractive drug target to treat norovirus infections. Genogroup II (GII) noroviruses are responsible for a majority of outbreaks; however, limited data exists regarding GII norovirus proteases. Methods: We report here successful expression, purification, characterization, and inhibition of the Minerva virus protease (MVpro), a genogroup II genotype 4 (GII.4) norovirus protease. We observed MVpro as both a monomer and dimer in solution through size-exclusion chromatography. In addition, MVpro cleaves the synthetic substrate mimicking the MVpro NS2/NS3 cleavage site more efficiently than other norovirus proteases such as the Norwalk virus protease (GI.1) and the MD145 protease (GII.4). Results and Conclusion: Compound A, a potent inhibitor of MVpro, is a good starting point for the design of inhibitors to target GII.4 noroviruses. Furthermore, the results presented here will allow for future characterization of MVpro inhibitors as they are synthesized.