来自密涅瓦病毒的GII.4诺如病毒蛋白酶的表达、纯化及特性分析
Expression, Purification and Characterization of a GII.4 Norovirus Protease from Minerva Virus.
作者信息
Kuiper Benjamin D, Muzzarelli Kendall M, Keusch Bradley J, Holcomb Joshua, Amblard Franck, Liu Peng, Zhou Shaoman, Kovari Iulia A, Yang Zhe, Schinazi Raymond F, Kovari Ladislau C
机构信息
Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, MI 48201, United States.
Center for AIDS Research, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, United States.
出版信息
Infect Disord Drug Targets. 2018;18(3):224-232. doi: 10.2174/1871526518666180521091158.
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.
背景
诺如病毒是全球急性胃肠炎的主要病因。负责切割病毒多聚蛋白的诺如病毒蛋白酶已成为治疗诺如病毒感染的一个有吸引力的药物靶点。II 基因组（GII）诺如病毒导致了大多数疫情爆发；然而，关于 GII 诺如病毒蛋白酶的数据有限。
方法
我们在此报告了 Minerva 病毒蛋白酶（MVpro）的成功表达、纯化、表征及抑制，MVpro 是一种 II 基因组 4 型（GII.4）诺如病毒蛋白酶。我们通过尺寸排阻色谱观察到 MVpro 在溶液中以单体和二聚体形式存在。此外，与其他诺如病毒蛋白酶如诺沃克病毒蛋白酶（GI.1）和 MD145 蛋白酶（GII.4）相比，MVpro 更有效地切割模拟 MVpro NS2/NS3 切割位点的合成底物。
结果与结论
化合物 A 是 MVpro 的一种有效抑制剂，是设计靶向 GII.4 诺如病毒的抑制剂的良好起点。此外，本文给出的结果将有助于在合成 MVpro 抑制剂时对其进行未来的表征。
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