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Title: A study on antigenicity and receptor-binding ability of fragment 450-650 of the spike protein of SARS coronavirus

Abstract

The spike (S) protein of SARS coronavirus (SARS-CoV) is responsible for viral binding with ACE2 molecules. Its receptor-binding motif (S-RBM) is located between residues 424 and 494, which folds into 2 anti-parallel {beta}-sheets, {beta}5 and {beta}6. We have previously demonstrated that fragment 450-650 of the S protein (S450-650) is predominantly recognized by convalescent sera of SARS patients. The N-terminal 60 residues (450-510) of the S450-650 fragment covers the entire {beta}6 strand of S-RBM. In the present study, we demonstrate that patient sera predominantly recognized 2 linear epitopes outside the {beta}6 fragment, while the mouse antisera, induced by immunization of BALB/c mice with recombinant S450-650, mainly recognized the {beta}6 strand-containing region. Unlike patient sera, however, the mouse antisera were unable to inhibit the infectivity of S protein-expressing (SARS-CoV-S) pseudovirus. Fusion protein between green fluorescence protein (GFP) and S450-650 (S450-650-GFP) was able to stain Vero E6 cells and deletion of the {beta}6 fragment rendered the fusion product (S511-650-GFP) unable to do so. Similarly, recombinant S450-650, but not S511-650, was able to block the infection of Vero E6 cells by the SARS-CoV-S pseudovirus. Co-precipitation experiments confirmed that S450-650 was able to specifically bind with ACE2 molecules in lysate of Vero E6 cells. However,more » the ability of S450-510, either alone or in fusion with GFP, to bind with ACE2 was significantly poorer compared with S450-650. Our data suggest a possibility that, although the {beta}6 strand alone is able to bind with ACE2 with relatively high affinity, residues outside the S-RBM could also assist the receptor binding of SARS-CoV-S protein.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2];  [1]
  1. Department of Immunology, Peking University Health Science Center, Peking University, 38 Xueyuan Road, Beijing 100083 (China)
  2. Institute of Basic Medical Sciences, Peking Union Medical College, Beijing (China)
Publication Date:
OSTI Identifier:
20976999
Resource Type:
Journal Article
Journal Name:
Virology
Additional Journal Information:
Journal Volume: 359; Journal Issue: 2; Other Information: DOI: 10.1016/j.virol.2006.09.022; PII: S0042-6822(06)00677-5; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0042-6822
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; COPRECIPITATION; FLUORESCENCE; INFECTIVITY; MICE; PATIENTS; RECEPTORS; RESIDUES; VIRAL DISEASES

Citation Formats

Jincun, Zhao, Wei, Wang, Zhihong, Yuan, Rujing, Jia, Zhendong, Zhao, Xiaojun, Xu, Ping, Lv, Yan, Zhang, Chengyu, Jiang, and Xiaoming, Gao. A study on antigenicity and receptor-binding ability of fragment 450-650 of the spike protein of SARS coronavirus. United States: N. p., 2007. Web. doi:10.1016/j.virol.2006.09.022.
Jincun, Zhao, Wei, Wang, Zhihong, Yuan, Rujing, Jia, Zhendong, Zhao, Xiaojun, Xu, Ping, Lv, Yan, Zhang, Chengyu, Jiang, & Xiaoming, Gao. A study on antigenicity and receptor-binding ability of fragment 450-650 of the spike protein of SARS coronavirus. United States. https://doi.org/10.1016/j.virol.2006.09.022
Jincun, Zhao, Wei, Wang, Zhihong, Yuan, Rujing, Jia, Zhendong, Zhao, Xiaojun, Xu, Ping, Lv, Yan, Zhang, Chengyu, Jiang, and Xiaoming, Gao. 2007. "A study on antigenicity and receptor-binding ability of fragment 450-650 of the spike protein of SARS coronavirus". United States. https://doi.org/10.1016/j.virol.2006.09.022.
@article{osti_20976999,
title = {A study on antigenicity and receptor-binding ability of fragment 450-650 of the spike protein of SARS coronavirus},
author = {Jincun, Zhao and Wei, Wang and Zhihong, Yuan and Rujing, Jia and Zhendong, Zhao and Xiaojun, Xu and Ping, Lv and Yan, Zhang and Chengyu, Jiang and Xiaoming, Gao},
abstractNote = {The spike (S) protein of SARS coronavirus (SARS-CoV) is responsible for viral binding with ACE2 molecules. Its receptor-binding motif (S-RBM) is located between residues 424 and 494, which folds into 2 anti-parallel {beta}-sheets, {beta}5 and {beta}6. We have previously demonstrated that fragment 450-650 of the S protein (S450-650) is predominantly recognized by convalescent sera of SARS patients. The N-terminal 60 residues (450-510) of the S450-650 fragment covers the entire {beta}6 strand of S-RBM. In the present study, we demonstrate that patient sera predominantly recognized 2 linear epitopes outside the {beta}6 fragment, while the mouse antisera, induced by immunization of BALB/c mice with recombinant S450-650, mainly recognized the {beta}6 strand-containing region. Unlike patient sera, however, the mouse antisera were unable to inhibit the infectivity of S protein-expressing (SARS-CoV-S) pseudovirus. Fusion protein between green fluorescence protein (GFP) and S450-650 (S450-650-GFP) was able to stain Vero E6 cells and deletion of the {beta}6 fragment rendered the fusion product (S511-650-GFP) unable to do so. Similarly, recombinant S450-650, but not S511-650, was able to block the infection of Vero E6 cells by the SARS-CoV-S pseudovirus. Co-precipitation experiments confirmed that S450-650 was able to specifically bind with ACE2 molecules in lysate of Vero E6 cells. However, the ability of S450-510, either alone or in fusion with GFP, to bind with ACE2 was significantly poorer compared with S450-650. Our data suggest a possibility that, although the {beta}6 strand alone is able to bind with ACE2 with relatively high affinity, residues outside the S-RBM could also assist the receptor binding of SARS-CoV-S protein.},
doi = {10.1016/j.virol.2006.09.022},
url = {https://www.osti.gov/biblio/20976999}, journal = {Virology},
issn = {0042-6822},
number = 2,
volume = 359,
place = {United States},
year = {Thu Mar 15 00:00:00 EDT 2007},
month = {Thu Mar 15 00:00:00 EDT 2007}
}