31nstitute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Taiyuan Road,Shanghai 200031, China.
31nstitute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Taiyuan Road,Shanghai 200031, China.
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发表时间：
2004
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通讯作者：
XiaodongWu;BoSHANG;RuiFuYANG;Haoyu;ZhiHalMA;Xushen;Yongyongji;Yinglin;YaDiWU;GuoMeiLIN
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作者：
XiaodongWu;BoSHANG;RuiFuYANG;Haoyu;ZhiHalMA;Xushen;Yongyongji;Yinglin;YaDiWU;GuoMeiLIN
刺突蛋白是SARS冠状病毒的主要结构蛋白之一。它对于病毒粒子与宿主细胞受体的相互作用以及随后病毒包膜与宿主细胞膜的融合以允许感染是必需的。一些冠状病毒的刺突蛋白，如MHV、HCoV-OC 43、AIBV和BcoV，被蛋白水解切割成两个亚基，S1和S2。相反，TGV，FIPV和HCoV-229 E不是。许多研究表明，纤突蛋白的切割严重影响其功能。为了研究SARS冠状病毒S蛋白的成熟和蛋白水解过程，我们制备了SARS冠状病毒S1和S2亚单位特异性抗体（Abs）以及N、E和3CL蛋白特异性抗体。Western blot结果表明，所制备的抗体能够与SARS-CoV感染Vero-E6细胞后的蛋白质特异性结合。电镜观察表明，抗S1和S2单克隆抗体能与SARSCoV结合。当用S2抗体与SARS-CoV感染细胞裂解液进行免疫沉淀时，用S2特异性mAb通过Western印迹分析检测到切割的S2片段。数据表明，在裂解物中观察到S蛋白的裂解，表明宿主细胞中存在S蛋白的蛋白水解加工。
Spike protein is one of the major structural proteins of severe acute respiratory syndrome-coronavirus. It is essential for the interaction of the virons with host cell receptors and subsequent fusion of the viral envelop with host cell membrane to allow infection. Some spike proteins of coronavirus, such as MHV, HCoV-OC43, AIBV and BcoV, are proteolytically cleaved into two subunits, S 1 and S2. In contrast, TGV, FIPV and HCoV-229E are not. Many studies have shown that the cleavage of spike protein seriously affects its function. In order to investigate the maturation and proteolytic processing of the S protein of SARS CoV, we generated S 1 and S2 subunit specific antibodies (Abs) as well as N, E and 3CL protein-specific Abs. Our results showed that the antibodies could efficiently and specifically bind to their corresponding proteins from E.coli expressed or lysate of SARS-CoV infected Vero-E6 cells by Western blot analysis. Furthermore, the anti-S 1 and S2 Abs were proved to be capable of binding to SARS CoV under electron microscope observation. When S2 Ab was used to perform immune precipitation with lysate of SARS-CoV infected cells, a cleaved S2 fragment was detected with S2-specific mAb by Western blot analysis. The data demonstrated that the cleavage of S protein was observed in the lysate, indicating that proteolytic processing of S protein is present in host cells.