Francesca R. Hills, Fátima Jorge, Laura N. Burga, Mihnea Bostina
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Abstract
Coronaviridae spike glycoproteins mediate viral entry and fusion to host cells through binding to host receptors (i.e., ACE2, DPP4) and are key components in determining viral host range, making them targets for antiviral research. Here, we describe the expression, purification, and characterization of recombinant spike proteins to aid in protein characterization and analysis. These protocols were used for the production of spike glycoproteins from civet, pangolin, and bat coronaviruses, as well as high-resolution cryo-electron microscopy (cryo-EM) structural analysis of bat and civet host coronavirus spike glycoproteins (Hills et al., 2024). © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC.
Basic Protocol 1 : Expression and purification of SARS-CoV spike protein from ExpiCHO cells
Basic Protocol 2 : Preparation of SARS-CoV spike protein for visualization by negative-stain transmission electron microscopy and cryo-electron microscopy
用于冷冻电镜分析的sars相关刺突糖蛋白的表达和纯化
冠状病毒刺突糖蛋白通过与宿主受体（如ACE2、DPP4）结合介导病毒进入和融合宿主细胞，是决定病毒宿主范围的关键成分，是抗病毒研究的靶点。本文描述了重组刺突蛋白的表达、纯化和表征，以帮助蛋白质表征和分析。这些方案用于从果子狸、穿山甲和蝙蝠冠状病毒中生产刺突糖蛋白，以及对蝙蝠和果子狸宿主冠状病毒刺突糖蛋白进行高分辨率冷冻电镜（cro - em）结构分析（Hills等，2024）。©2025作者。当前方案发表于Wiley期刊有限责任公司。基本方案1：从ExpiCHO细胞中表达和纯化SARS-CoV刺状蛋白基本方案2：制备SARS-CoV刺状蛋白用于负染色透射电镜和冷冻电镜的可视化
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