A novel cell-based binding assay system reconstituting interaction between SARS-CoV S protein and its cellular receptor
Identifieur interne : 005489 ( Main/Curation ); précédent : 005488; suivant : 005490A novel cell-based binding assay system reconstituting interaction between SARS-CoV S protein and its cellular receptor
Auteurs : Chih-Fong Chou ; Shuo Shen ; Yee-Joo Tan ; Burtram C. Fielding ; Timothy H. P. Tan ; Jianlin Fu ; Qiurong Xu ; Seng Gee Lim ; Wanjin HongSource :
- Journal of Virological Methods [ 0166-0934 ] ; 2004.
Descripteurs français
- KwdFr :
- Animaux, Cellules CHO, Cellules Vero, Cricetinae, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (métabolisme), Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (métabolisme), Protéines à fluorescence verte (génétique), Protéines à fluorescence verte (métabolisme), Récepteurs viraux (métabolisme), Virus du SRAS (métabolisme).
- MESH :
- génétique : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines à fluorescence verte.
- métabolisme : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines à fluorescence verte, Récepteurs viraux, Virus du SRAS.
- Animaux, Cellules CHO, Cellules Vero, Cricetinae, Glycoprotéine de spicule des coronavirus.
English descriptors
- KwdEn :
- Animals, CHO Cells, Chlorocebus aethiops, Cricetinae, Green Fluorescent Proteins (genetics), Green Fluorescent Proteins (metabolism), Membrane Glycoproteins (genetics), Membrane Glycoproteins (metabolism), Receptors, Virus (metabolism), SARS Virus (metabolism), Spike Glycoprotein, Coronavirus, Vero Cells, Viral Envelope Proteins (genetics), Viral Envelope Proteins (metabolism).
- MESH :
- chemical , genetics : Green Fluorescent Proteins, Membrane Glycoproteins, Viral Envelope Proteins.
- chemical , metabolism : Green Fluorescent Proteins, Membrane Glycoproteins, Receptors, Virus, Viral Envelope Proteins.
- metabolism : SARS Virus.
- Animals, CHO Cells, Chlorocebus aethiops, Cricetinae, Spike Glycoprotein, Coronavirus, Vero Cells.
Abstract
Severe acute respiratory syndrome (SARS), a life-threatening disease, is caused by the newly identified virus SARS coronavirus (SARS-CoV). In order to study the spike (S) protein of this highly contagious virus, we established a clonal cell-line, CHO-SG, from the Chinese hamster ovary cells that stably expresses C-terminally EGFP-tagged SARS-CoV S protein (S-EGFP). The ectodomain of the S glycoprotein is localized on the surface of CHO-SG cells with
Url:
DOI: 10.1016/j.jviromet.2004.09.008
PubMed: 15582697
PubMed Central: 7112911
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PMC:7112911Le document en format XML
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<author><name sortKey="Chou, Chih Fong" sort="Chou, Chih Fong" uniqKey="Chou C" first="Chih-Fong" last="Chou">Chih-Fong Chou</name>
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<author><name sortKey="Shen, Shuo" sort="Shen, Shuo" uniqKey="Shen S" first="Shuo" last="Shen">Shuo Shen</name>
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<author><name sortKey="Tan, Yee Joo" sort="Tan, Yee Joo" uniqKey="Tan Y" first="Yee-Joo" last="Tan">Yee-Joo Tan</name>
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<author><name sortKey="Fielding, Burtram C" sort="Fielding, Burtram C" uniqKey="Fielding B" first="Burtram C." last="Fielding">Burtram C. Fielding</name>
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<author><name sortKey="Tan, Timothy H P" sort="Tan, Timothy H P" uniqKey="Tan T" first="Timothy H. P." last="Tan">Timothy H. P. Tan</name>
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<author><name sortKey="Fu, Jianlin" sort="Fu, Jianlin" uniqKey="Fu J" first="Jianlin" last="Fu">Jianlin Fu</name>
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<author><name sortKey="Xu, Qiurong" sort="Xu, Qiurong" uniqKey="Xu Q" first="Qiurong" last="Xu">Qiurong Xu</name>
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<author><name sortKey="Lim, Seng Gee" sort="Lim, Seng Gee" uniqKey="Lim S" first="Seng Gee" last="Lim">Seng Gee Lim</name>
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<author><name sortKey="Hong, Wanjin" sort="Hong, Wanjin" uniqKey="Hong W" first="Wanjin" last="Hong">Wanjin Hong</name>
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<series><title level="j">Journal of Virological Methods</title>
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<term>CHO Cells</term>
<term>Chlorocebus aethiops</term>
<term>Cricetinae</term>
<term>Green Fluorescent Proteins (genetics)</term>
<term>Green Fluorescent Proteins (metabolism)</term>
<term>Membrane Glycoproteins (genetics)</term>
<term>Membrane Glycoproteins (metabolism)</term>
<term>Receptors, Virus (metabolism)</term>
<term>SARS Virus (metabolism)</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Vero Cells</term>
<term>Viral Envelope Proteins (genetics)</term>
<term>Viral Envelope Proteins (metabolism)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term>
<term>Cellules CHO</term>
<term>Cellules Vero</term>
<term>Cricetinae</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires (génétique)</term>
<term>Glycoprotéines membranaires (métabolisme)</term>
<term>Protéines de l'enveloppe virale (génétique)</term>
<term>Protéines de l'enveloppe virale (métabolisme)</term>
<term>Protéines à fluorescence verte (génétique)</term>
<term>Protéines à fluorescence verte (métabolisme)</term>
<term>Récepteurs viraux (métabolisme)</term>
<term>Virus du SRAS (métabolisme)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Green Fluorescent Proteins</term>
<term>Membrane Glycoproteins</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Green Fluorescent Proteins</term>
<term>Membrane Glycoproteins</term>
<term>Receptors, Virus</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines à fluorescence verte</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines à fluorescence verte</term>
<term>Récepteurs viraux</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>CHO Cells</term>
<term>Chlorocebus aethiops</term>
<term>Cricetinae</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Vero Cells</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Cellules CHO</term>
<term>Cellules Vero</term>
<term>Cricetinae</term>
<term>Glycoprotéine de spicule des coronavirus</term>
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<front><div type="abstract" xml:lang="en"><p>Severe acute respiratory syndrome (SARS), a life-threatening disease, is caused by the newly identified virus SARS coronavirus (SARS-CoV). In order to study the spike (S) protein of this highly contagious virus, we established a clonal cell-line, CHO-SG, from the Chinese hamster ovary cells that stably expresses C-terminally EGFP-tagged SARS-CoV S protein (S-EGFP). The ectodomain of the S glycoprotein is localized on the surface of CHO-SG cells with <italic>N</italic>
-acetyl-glucosamine-terminated carbohydrate structure. CHO-SG cells associated tightly with Vero E6 cells, a SARS-CoV receptor (ACE2) expressing cell-line, and the interaction remained stable under highly stringent condition (1 M NaCl). This interaction could be blocked by either the serum from a SARS convalescent patient or a goat anti-ACE2 antibody, indicating that the interaction is specific. A binding epitope with lesser degree of glycosylation and native conformation was localized by using rabbit anti-sera raised against five denatured recombinant S protein fragments expressed in <italic>Escherichia coli</italic>
. One of the sera obtained from the fragment encompassing amino acids 48-358 significantly blocked the interaction between CHO-SG and Vero E6 cells. The region is useful for studying neutralizing antibodies in future vaccine development. This paper describes an easy and safe cell-based assay suitable for studying the binding between SARS-CoV S protein and its receptor.</p>
</div>
</front>
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</TEI>
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