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Expression, Purification and Characterization of a GII.4 Norovirus Protease from Minerva Virus

[ Vol. 18 , Issue. 3 ]

Author(s):

Benjamin D. Kuiper, Kendall M. Muzzarelli, Bradley J. Keusch, Joshua Holcomb, Franck Amblard, Peng Liu, Shaoman Zhou, Iulia A. Kovari, Zhe Yang, Raymond F. Schinazi and Ladislau C. Kovari*   Pages 224 - 232 ( 9 )

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 sizeexclusion 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.

Keywords:

Noroviruses, Polyprotein, Chromatography, Potent inhibitor, MVpro inhibitors, Purification.

Affiliation:

Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201

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