Characterization and inhibition of norovirus proteases of genogroups I and II using a fluorescence resonance energy transfer assay

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• 作者：Kyeong-Ok Chang^a ; Daisuke Takahashi^b ; Om Prakash^b ; Yunjeong Kim^a ; ^{ykim@vet.ksu.edu}
• 关键词：Antiviral ; Viral proteases ; Fluorescence resonance energy transfer ; Norwalk virus ; MD145 virus ; Nuclear magnetic resonance spectroscopy ; Chymostatin
• 刊名：Virology
• 出版年：2012
• 出版时间：20 February 2012
• 年：2012
• 卷：423
• 期：2
• 页码：125-133
• 全文大小：1520 K

文摘

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.