Volume 14, No 5, Oct 2004
ISSN: 1001-0602
EISSN: 1748-7838 2018
impact factor 17.848*
(Clarivate Analytics, 2019)
Volume 14 Issue 5, October 2004: 400-406
ORIGINAL ARTICLES
The spike protein of severe acute respiratory syndrome (SARS) is cleaved in virus infected Vero-E6 cells
Xiao Dong WU 1*, Bo Shang 1*, Rui Fu YANG2*, Hao YU3*, Zhi Hai Ma1*, Xu SHEN3*, Yong Yong JI1, Ying LIN 1, Ya Di Wu1, Guo Mei Lin1, Lin TIAN1, Xiao Qing GAN1, Sheng YANG2, Wei Hong JIANG2, Er Hei DAI4, Xiao Yi WANG4, Hua Liang JIANG3, You Hua Xie1, Xue Liang Zhu1, Gang PEI1, Lin LI1, Jia Rui WU1**, Bing SUN 1**
1Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.
2Institute of Microbiology and Epidemiology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100071, China.
3Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200031, China.
4Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Taiyuan Road, Shanghai 200031, China.
Correspondence: Jia Rui WU Bing SUN(wujr@sibs.ac.cn bsun@sibs.ac.cn)
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, S1 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 S1 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-S1 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.
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