Guo Jingxu, Douangamath Alice, Song Weixiao, Coker Alun R, Chan A W Edith, Wood Steve P, Cooper Jonathan B, Resnick Efrat, London Nir, Delft Frank von
Division of Medicine, UCL, Gower Street, London WC1E 6BT, UK.
Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE, UK.
J Struct Biol X. 2020 Jul 16;4:100031. doi: 10.1016/j.yjsbx.2020.100031. eCollection 2020.
Outbreaks of human epidemic nonbacterial gastroenteritis are mainly caused by noroviruses. Viral replication requires a 3C-like cysteine protease (3CL) which processes the 200 kDa viral polyprotein into six functional proteins. The 3CL has attracted much interest due to its potential as a target for antiviral drugs. A system for growing high-quality crystals of native Southampton norovirus 3CL (SV3CP) has been established, allowing the ligand-free crystal structure to be determined to 1.3 Å in a tetrameric state. This also allowed crystal-based fragment screening to be performed with various compound libraries, ultimately to guide drug discovery for SV3CP. A total of 19 fragments were found to bind to the protease out of the 844 which were screened. Two of the hits were located at the active site of SV3CP and showed good inhibitory activity in kinetic assays. Another 5 were found at the enzyme's putative RNA-binding site and a further 11 were located in the symmetric central cavity of the tetramer.
人类流行性非细菌性肠胃炎的爆发主要由诺如病毒引起。病毒复制需要一种类3C半胱氨酸蛋白酶（3CL），它将200 kDa的病毒多聚蛋白加工成六种功能蛋白。3CL因其作为抗病毒药物靶点的潜力而备受关注。已建立了一种用于培养南安普敦诺如病毒天然3CL（SV3CP）高质量晶体的系统，使得能够在四聚体状态下将无配体晶体结构解析到1.3 Å。这也使得能够使用各种化合物库进行基于晶体的片段筛选，最终指导针对SV3CP的药物发现。在筛选的844个片段中，共发现19个片段与该蛋白酶结合。其中两个命中片段位于SV3CP的活性位点，在动力学分析中显示出良好的抑制活性。另外5个在该酶假定的RNA结合位点被发现，还有11个位于四聚体的对称中心腔中。
