The spike protein of severe acute respiratory syndrome (SARS) is cleaved in virus infected Vero-E6 cells

Xiao Dong Wu, Bo Shang, Rui Fu Yang, Hao Yu, Zhi Hai Ma, Xu Shen, Yong Yong Ji, Ying Lin, Ya Di Wu, Guo Mei Lin, Lin Tian, Xiao Qing Gan, Sheng Yang, Wei Hong Jiang, Er Hei Dai, Xiao Yi Wang, Hua Liang Jiang, You Hua Xie, Xue Liang Zhu, Gang PeiLin Li, Jia Rui Wu, Bing Sun

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)400-406
Number of pages7
JournalCell Research
Volume14
Issue number5
DOIs
StatePublished - Oct 2004

Keywords

  • 3CL protease
  • Envelope protein
  • Nucleocapsid protein
  • Polyclonal antibody
  • SARS-CoV
  • Spike protein

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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