A study on antigenicity and receptor-binding ability of fragment 450-650 of the spike protein of SARS coronavirus.
Identifieur interne : 003775 ( Main/Exploration ); précédent : 003774; suivant : 003776A study on antigenicity and receptor-binding ability of fragment 450-650 of the spike protein of SARS coronavirus.
Auteurs : Jincun Zhao [République populaire de Chine] ; Wei Wang ; Zhihong Yuan ; Rujing Jia ; Zhendong Zhao ; Xiaojun Xu ; Ping Lv ; Yan Zhang ; Chengyu Jiang ; Xiao-Ming GaoSource :
- Virology [ 0042-6822 ] ; 2007.
Descripteurs français
- KwdFr :
- Affinité des anticorps, Animaux, Anticorps antiviraux, Cellules Vero, Conformation des protéines, Déterminants antigéniques des lymphocytes B, Femelle, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (immunologie), Glycoprotéines membranaires (métabolisme), Humains, Lignée cellulaire, Modèles moléculaires, Peptidyl-Dipeptidase A (métabolisme), Protéines de l'enveloppe virale (immunologie), Protéines de l'enveloppe virale (métabolisme), Récepteurs viraux (métabolisme), Régulation de l'expression des gènes viraux, Souris, Souris de lignée BALB C, Virus du SRAS (immunologie), Virus du SRAS (métabolisme).
- MESH :
- immunologie : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Virus du SRAS.
- métabolisme : Glycoprotéines membranaires, Peptidyl-Dipeptidase A, Protéines de l'enveloppe virale, Récepteurs viraux, Virus du SRAS.
- Affinité des anticorps, Animaux, Anticorps antiviraux, Cellules Vero, Conformation des protéines, Déterminants antigéniques des lymphocytes B, Femelle, Glycoprotéine de spicule des coronavirus, Humains, Lignée cellulaire, Modèles moléculaires, Régulation de l'expression des gènes viraux, Souris, Souris de lignée BALB C.
English descriptors
- KwdEn :
- Animals, Antibodies, Viral, Antibody Affinity, Cell Line, Chlorocebus aethiops, Epitopes, B-Lymphocyte, Female, Gene Expression Regulation, Viral, Humans, Membrane Glycoproteins (immunology), Membrane Glycoproteins (metabolism), Mice, Mice, Inbred BALB C, Models, Molecular, Peptidyl-Dipeptidase A (metabolism), Protein Conformation, Receptors, Virus (metabolism), SARS Virus (immunology), SARS Virus (metabolism), Spike Glycoprotein, Coronavirus, Vero Cells, Viral Envelope Proteins (immunology), Viral Envelope Proteins (metabolism).
- MESH :
- chemical , immunology : Membrane Glycoproteins, Viral Envelope Proteins.
- chemical , metabolism : Membrane Glycoproteins, Peptidyl-Dipeptidase A, Receptors, Virus, Viral Envelope Proteins.
- chemical : Antibodies, Viral, Epitopes, B-Lymphocyte, Spike Glycoprotein, Coronavirus.
- immunology : SARS Virus.
- metabolism : SARS Virus.
- Animals, Antibody Affinity, Cell Line, Chlorocebus aethiops, Female, Gene Expression Regulation, Viral, Humans, Mice, Mice, Inbred BALB C, Models, Molecular, Protein Conformation, Vero Cells.
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, beta5 and beta6. 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 beta6 strand of S-RBM. In the present study, we demonstrate that patient sera predominantly recognized 2 linear epitopes outside the beta6 fragment, while the mouse antisera, induced by immunization of BALB/c mice with recombinant S450-650, mainly recognized the beta6 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 beta6 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 beta6 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
PubMed: 17055551
Affiliations:
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Zhihong" sort="Yuan, Zhihong" uniqKey="Yuan Z" first="Zhihong" last="Yuan">Zhihong Yuan</name></author><author><name sortKey="Jia, Rujing" sort="Jia, Rujing" uniqKey="Jia R" first="Rujing" last="Jia">Rujing Jia</name></author><author><name sortKey="Zhao, Zhendong" sort="Zhao, Zhendong" uniqKey="Zhao Z" first="Zhendong" last="Zhao">Zhendong Zhao</name></author><author><name sortKey="Xu, Xiaojun" sort="Xu, Xiaojun" uniqKey="Xu X" first="Xiaojun" last="Xu">Xiaojun Xu</name></author><author><name sortKey="Lv, Ping" sort="Lv, Ping" uniqKey="Lv P" first="Ping" last="Lv">Ping Lv</name></author><author><name sortKey="Zhang, Yan" sort="Zhang, Yan" uniqKey="Zhang Y" first="Yan" last="Zhang">Yan Zhang</name></author><author><name sortKey="Jiang, Chengyu" sort="Jiang, Chengyu" uniqKey="Jiang C" first="Chengyu" last="Jiang">Chengyu Jiang</name></author><author><name sortKey="Gao, Xiao Ming" sort="Gao, Xiao Ming" uniqKey="Gao X" first="Xiao-Ming" last="Gao">Xiao-Ming Gao</name></author></analytic><series><title level="j">Virology</title><idno type="ISSN">0042-6822</idno><imprint><date when="2007" type="published">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Viral</term><term>Antibody Affinity</term><term>Cell Line</term><term>Chlorocebus aethiops</term><term>Epitopes, B-Lymphocyte</term><term>Female</term><term>Gene Expression Regulation, Viral</term><term>Humans</term><term>Membrane Glycoproteins (immunology)</term><term>Membrane Glycoproteins (metabolism)</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Models, Molecular</term><term>Peptidyl-Dipeptidase A (metabolism)</term><term>Protein Conformation</term><term>Receptors, Virus (metabolism)</term><term>SARS Virus (immunology)</term><term>SARS Virus (metabolism)</term><term>Spike Glycoprotein, Coronavirus</term><term>Vero Cells</term><term>Viral Envelope Proteins (immunology)</term><term>Viral Envelope Proteins (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Affinité des anticorps</term><term>Animaux</term><term>Anticorps antiviraux</term><term>Cellules Vero</term><term>Conformation des protéines</term><term>Déterminants antigéniques des lymphocytes B</term><term>Femelle</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (immunologie)</term><term>Glycoprotéines membranaires (métabolisme)</term><term>Humains</term><term>Lignée cellulaire</term><term>Modèles moléculaires</term><term>Peptidyl-Dipeptidase A (métabolisme)</term><term>Protéines de l'enveloppe virale (immunologie)</term><term>Protéines de l'enveloppe virale (métabolisme)</term><term>Récepteurs viraux (métabolisme)</term><term>Régulation de l'expression des gènes viraux</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Virus du SRAS (immunologie)</term><term>Virus du SRAS (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Membrane Glycoproteins</term><term>Peptidyl-Dipeptidase A</term><term>Receptors, Virus</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Antibodies, Viral</term><term>Epitopes, B-Lymphocyte</term><term>Spike Glycoprotein, Coronavirus</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</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>Peptidyl-Dipeptidase A</term><term>Protéines de l'enveloppe virale</term><term>Récepteurs viraux</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Antibody Affinity</term><term>Cell Line</term><term>Chlorocebus aethiops</term><term>Female</term><term>Gene Expression Regulation, Viral</term><term>Humans</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Models, Molecular</term><term>Protein Conformation</term><term>Vero Cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Affinité des anticorps</term><term>Animaux</term><term>Anticorps antiviraux</term><term>Cellules Vero</term><term>Conformation des protéines</term><term>Déterminants antigéniques des lymphocytes B</term><term>Femelle</term><term>Glycoprotéine de spicule des coronavirus</term><term>Humains</term><term>Lignée cellulaire</term><term>Modèles moléculaires</term><term>Régulation de l'expression des gènes viraux</term><term>Souris</term><term>Souris de lignée BALB C</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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, beta5 and beta6. 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 beta6 strand of S-RBM. In the present study, we demonstrate that patient sera predominantly recognized 2 linear epitopes outside the beta6 fragment, while the mouse antisera, induced by immunization of BALB/c mice with recombinant S450-650, mainly recognized the beta6 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 beta6 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 beta6 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.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Gao, Xiao Ming" sort="Gao, Xiao Ming" uniqKey="Gao X" first="Xiao-Ming" last="Gao">Xiao-Ming Gao</name><name sortKey="Jia, Rujing" sort="Jia, Rujing" uniqKey="Jia R" first="Rujing" last="Jia">Rujing Jia</name><name sortKey="Jiang, Chengyu" sort="Jiang, Chengyu" uniqKey="Jiang C" first="Chengyu" last="Jiang">Chengyu Jiang</name><name sortKey="Lv, Ping" sort="Lv, Ping" uniqKey="Lv P" first="Ping" last="Lv">Ping Lv</name><name sortKey="Wang, Wei" sort="Wang, Wei" uniqKey="Wang W" first="Wei" last="Wang">Wei Wang</name><name sortKey="Xu, Xiaojun" sort="Xu, Xiaojun" uniqKey="Xu X" first="Xiaojun" last="Xu">Xiaojun Xu</name><name sortKey="Yuan, Zhihong" sort="Yuan, Zhihong" uniqKey="Yuan Z" first="Zhihong" last="Yuan">Zhihong Yuan</name><name sortKey="Zhang, Yan" sort="Zhang, Yan" uniqKey="Zhang Y" first="Yan" last="Zhang">Yan Zhang</name><name sortKey="Zhao, Zhendong" sort="Zhao, Zhendong" uniqKey="Zhao Z" first="Zhendong" last="Zhao">Zhendong Zhao</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Zhao, Jincun" sort="Zhao, Jincun" uniqKey="Zhao J" first="Jincun" last="Zhao">Jincun Zhao</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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