Characterization and inhibition of norovirus proteases of genogroups I and II using a fluorescence resonance energy transfer assay.
Noroviruses are the major cause of food- or water-borne gastroenteritis outbreaks in humans. The norovirus protease that cleaves a large viral polyprotein to nonstructural proteins is essential for virus replication and an attractive target for antiviral drug development. Noroviruses show high genetic diversity with at least five genogroups, GI-GV, of which GI and GII are responsible for the majority of norovirus infections in humans. We cloned and expressed proteases of Norwalk virus (GI) and MD145 virus (GII) and characterized the enzymatic activities with fluorescence resonance energy transfer substrates. We demonstrated that the GI and GII proteases cleaved the substrates derived from the naturally occurring cleavage site in the open reading frame (ORF) 1 of G1 norovirus with similar efficiency, and that enzymatic activity of both proteases was inhibited by commercial protease inhibitors including chymostatin. The interaction of chymostatin to Norwalk virus protease was validated by nuclear magnetic resonance (NMR) spectroscopy.
Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, KS 66506, USA.
Kyeong-Ok
KO
Chang
Daisuke
D
Takahashi
Om
O
Prakash
Yunjeong
Y
Kim
D000595:Amino Acid Sequence; D015536:Down-Regulation; D031541:Fluorescence Resonance Energy Transfer; D005759:Gastroenteritis / Q000821:virology*; D005838:Genotype; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D029322:Norovirus / Q000737:chemistry / Q000145:classification / Q000201:enzymology / Q000235:genetics; D009663:Norwalk virus / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011480:Protease Inhibitors / Q000737:chemistry / Q000494:pharmacology; D016415:Sequence Alignment; D014764:Viral Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D009842:Oligopeptides; D011480:Protease Inhibitors; D014764:Viral Proteins; C002101:chymostatin; D010447:Peptide Hydrolases