Antigenicity and receptor-binding ability of recombinant SARS coronavirus spike protein.
Identifieur interne : 005457 ( Main/Exploration ); précédent : 005456; suivant : 005458Antigenicity and receptor-binding ability of recombinant SARS coronavirus spike protein.
Auteurs : Tin-Yun Ho [Taïwan] ; Shih-Lu Wu ; Shin-Ei Cheng ; Yen-Chiao Wei ; Shan-Ping Huang ; Chien-Yun HsiangSource :
- Biochemical and biophysical research communications [ 0006-291X ] ; 2004.
Descripteurs français
- KwdFr :
- ADN viral (génétique), Animaux, Antigènes viraux (génétique), Cellules Vero, Clonage moléculaire, Données de séquences moléculaires, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (), Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (immunologie), Glycoprotéines membranaires (métabolisme), Humains, Protéines de l'enveloppe virale (), Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (immunologie), Protéines de l'enveloppe virale (métabolisme), Protéines recombinantes (), Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Protéines recombinantes (métabolisme), Récepteurs viraux (métabolisme), Similitude de séquences d'acides aminés, Structure secondaire des protéines, Séquence d'acides aminés, Séquence nucléotidique, Virus du SRAS (génétique), Virus du SRAS (immunologie), Virus du SRAS (métabolisme), Virus du SRAS (pathogénicité).
- MESH :
- génétique : ADN viral, Antigènes viraux, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines recombinantes, Virus du SRAS.
- immunologie : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines recombinantes, Virus du SRAS.
- métabolisme : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines recombinantes, Récepteurs viraux, Virus du SRAS.
- pathogénicité : Virus du SRAS.
- Animaux, Cellules Vero, Clonage moléculaire, Données de séquences moléculaires, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires, Humains, Protéines de l'enveloppe virale, Protéines recombinantes, Similitude de séquences d'acides aminés, Structure secondaire des protéines, Séquence d'acides aminés, Séquence nucléotidique.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, Antigens, Viral (genetics), Base Sequence, Chlorocebus aethiops, Cloning, Molecular, DNA, Viral (genetics), Humans, Membrane Glycoproteins (chemistry), Membrane Glycoproteins (genetics), Membrane Glycoproteins (immunology), Membrane Glycoproteins (metabolism), Molecular Sequence Data, Protein Structure, Secondary, Receptors, Virus (metabolism), Recombinant Proteins (chemistry), Recombinant Proteins (genetics), Recombinant Proteins (immunology), Recombinant Proteins (metabolism), SARS Virus (genetics), SARS Virus (immunology), SARS Virus (metabolism), SARS Virus (pathogenicity), Sequence Homology, Amino Acid, Spike Glycoprotein, Coronavirus, Vero Cells, Viral Envelope Proteins (chemistry), Viral Envelope Proteins (genetics), Viral Envelope Proteins (immunology), Viral Envelope Proteins (metabolism).
- MESH :
- chemical , chemistry : Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , genetics : Antigens, Viral, DNA, Viral, Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , immunology : Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , metabolism : Membrane Glycoproteins, Receptors, Virus, Recombinant Proteins, Viral Envelope Proteins.
- genetics : SARS Virus.
- immunology : SARS Virus.
- metabolism : SARS Virus.
- pathogenicity : SARS Virus.
- Amino Acid Sequence, Animals, Base Sequence, Chlorocebus aethiops, Cloning, Molecular, Humans, Molecular Sequence Data, Protein Structure, Secondary, Sequence Homology, Amino Acid, Spike Glycoprotein, Coronavirus, Vero Cells.
Abstract
Severe acute respiratory syndrome (SARS) is an emerging infectious disease associated with a novel coronavirus and causing worldwide outbreaks. SARS coronavirus (SARS-CoV) is an enveloped RNA virus, which contains several structural proteins. Among these proteins, spike (S) protein is responsible for binding to specific cellular receptors and is a major antigenic determinant, which induces neutralizing antibody. In order to analyze the antigenicity and receptor-binding ability of SARS-CoV S protein, we expressed the S protein in Escherichia coli using a pET expression vector. After the isopropyl-beta-D-thiogalactoside induction, S protein was expressed in the soluble form and purified by nickel-affinity chromatography to homogeneity. The amount of S protein recovered was 0.2-0.3mg/100ml bacterial culture. The S protein was recognized by sera from SARS patients by ELISA and Western blot, which indicated that recombinant S protein retained its antigenicity. By biotinylated ELISA and Western blot using biotin-labeled S protein as the probe, we identified 130-kDa and 140-kDa proteins in Vero cells that might be the cellular receptors responsible for SARS-CoV infection. Taken together, these results suggested that recombinant S protein exhibited the antigenicity and receptor-binding ability, and it could be a good candidate for further developing SARS vaccine and anti-SARS therapy.
DOI: 10.1016/j.bbrc.2003.11.180
PubMed: 14706633
Affiliations:
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Le document en format XML
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last="Wei">Yen-Chiao Wei</name></author><author><name sortKey="Huang, Shan Ping" sort="Huang, Shan Ping" uniqKey="Huang S" first="Shan-Ping" last="Huang">Shan-Ping Huang</name></author><author><name sortKey="Hsiang, Chien Yun" sort="Hsiang, Chien Yun" uniqKey="Hsiang C" first="Chien-Yun" last="Hsiang">Chien-Yun Hsiang</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2004">2004</date><idno type="RBID">pubmed:14706633</idno><idno type="pmid">14706633</idno><idno type="doi">10.1016/j.bbrc.2003.11.180</idno><idno type="wicri:Area/PubMed/Corpus">003013</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">003013</idno><idno type="wicri:Area/PubMed/Curation">003013</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">003013</idno><idno type="wicri:Area/PubMed/Checkpoint">002E21</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002E21</idno><idno 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Taichung</wicri:regionArea><wicri:noRegion>Taichung</wicri:noRegion></affiliation></author><author><name sortKey="Wu, Shih Lu" sort="Wu, Shih Lu" uniqKey="Wu S" first="Shih-Lu" last="Wu">Shih-Lu Wu</name></author><author><name sortKey="Cheng, Shin Ei" sort="Cheng, Shin Ei" uniqKey="Cheng S" first="Shin-Ei" last="Cheng">Shin-Ei Cheng</name></author><author><name sortKey="Wei, Yen Chiao" sort="Wei, Yen Chiao" uniqKey="Wei Y" first="Yen-Chiao" last="Wei">Yen-Chiao Wei</name></author><author><name sortKey="Huang, Shan Ping" sort="Huang, Shan Ping" uniqKey="Huang S" first="Shan-Ping" last="Huang">Shan-Ping Huang</name></author><author><name sortKey="Hsiang, Chien Yun" sort="Hsiang, Chien Yun" uniqKey="Hsiang C" first="Chien-Yun" last="Hsiang">Chien-Yun Hsiang</name></author></analytic><series><title level="j">Biochemical and biophysical research communications</title><idno type="ISSN">0006-291X</idno><imprint><date when="2004" type="published">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Antigens, Viral (genetics)</term><term>Base Sequence</term><term>Chlorocebus aethiops</term><term>Cloning, Molecular</term><term>DNA, Viral (genetics)</term><term>Humans</term><term>Membrane Glycoproteins (chemistry)</term><term>Membrane Glycoproteins (genetics)</term><term>Membrane Glycoproteins (immunology)</term><term>Membrane Glycoproteins (metabolism)</term><term>Molecular Sequence Data</term><term>Protein Structure, Secondary</term><term>Receptors, Virus (metabolism)</term><term>Recombinant Proteins (chemistry)</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (immunology)</term><term>Recombinant Proteins (metabolism)</term><term>SARS Virus (genetics)</term><term>SARS Virus (immunology)</term><term>SARS Virus (metabolism)</term><term>SARS Virus (pathogenicity)</term><term>Sequence Homology, Amino Acid</term><term>Spike Glycoprotein, Coronavirus</term><term>Vero Cells</term><term>Viral Envelope Proteins (chemistry)</term><term>Viral Envelope Proteins (genetics)</term><term>Viral Envelope Proteins (immunology)</term><term>Viral Envelope Proteins (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN viral (génétique)</term><term>Animaux</term><term>Antigènes viraux (génétique)</term><term>Cellules Vero</term><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires ()</term><term>Glycoprotéines membranaires (génétique)</term><term>Glycoprotéines membranaires (immunologie)</term><term>Glycoprotéines membranaires (métabolisme)</term><term>Humains</term><term>Protéines de l'enveloppe virale ()</term><term>Protéines de l'enveloppe virale (génétique)</term><term>Protéines de l'enveloppe virale (immunologie)</term><term>Protéines de l'enveloppe virale (métabolisme)</term><term>Protéines recombinantes ()</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (immunologie)</term><term>Protéines recombinantes (métabolisme)</term><term>Récepteurs viraux (métabolisme)</term><term>Similitude de séquences d'acides aminés</term><term>Structure secondaire des protéines</term><term>Séquence d'acides aminés</term><term>Séquence nucléotidique</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (immunologie)</term><term>Virus du SRAS (métabolisme)</term><term>Virus du SRAS (pathogénicité)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Membrane Glycoproteins</term><term>Recombinant Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antigens, Viral</term><term>DNA, Viral</term><term>Membrane Glycoproteins</term><term>Recombinant Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Membrane Glycoproteins</term><term>Recombinant Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Membrane Glycoproteins</term><term>Receptors, Virus</term><term>Recombinant Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN viral</term><term>Antigènes viraux</term><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Protéines recombinantes</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Protéines recombinantes</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>Protéines de l'enveloppe virale</term><term>Protéines recombinantes</term><term>Récepteurs viraux</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="pathogenicity" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="pathogénicité" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Base Sequence</term><term>Chlorocebus aethiops</term><term>Cloning, Molecular</term><term>Humans</term><term>Molecular Sequence Data</term><term>Protein Structure, Secondary</term><term>Sequence Homology, Amino Acid</term><term>Spike Glycoprotein, Coronavirus</term><term>Vero Cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules Vero</term><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires</term><term>Humains</term><term>Protéines de l'enveloppe virale</term><term>Protéines recombinantes</term><term>Similitude de séquences d'acides aminés</term><term>Structure secondaire des protéines</term><term>Séquence d'acides aminés</term><term>Séquence nucléotidique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) is an emerging infectious disease associated with a novel coronavirus and causing worldwide outbreaks. SARS coronavirus (SARS-CoV) is an enveloped RNA virus, which contains several structural proteins. Among these proteins, spike (S) protein is responsible for binding to specific cellular receptors and is a major antigenic determinant, which induces neutralizing antibody. In order to analyze the antigenicity and receptor-binding ability of SARS-CoV S protein, we expressed the S protein in Escherichia coli using a pET expression vector. After the isopropyl-beta-D-thiogalactoside induction, S protein was expressed in the soluble form and purified by nickel-affinity chromatography to homogeneity. The amount of S protein recovered was 0.2-0.3mg/100ml bacterial culture. The S protein was recognized by sera from SARS patients by ELISA and Western blot, which indicated that recombinant S protein retained its antigenicity. By biotinylated ELISA and Western blot using biotin-labeled S protein as the probe, we identified 130-kDa and 140-kDa proteins in Vero cells that might be the cellular receptors responsible for SARS-CoV infection. Taken together, these results suggested that recombinant S protein exhibited the antigenicity and receptor-binding ability, and it could be a good candidate for further developing SARS vaccine and anti-SARS therapy.</div></front></TEI><affiliations><list><country><li>Taïwan</li></country></list><tree><noCountry><name sortKey="Cheng, Shin Ei" sort="Cheng, Shin Ei" uniqKey="Cheng S" first="Shin-Ei" last="Cheng">Shin-Ei Cheng</name><name sortKey="Hsiang, Chien Yun" sort="Hsiang, Chien Yun" uniqKey="Hsiang C" first="Chien-Yun" last="Hsiang">Chien-Yun Hsiang</name><name sortKey="Huang, Shan Ping" sort="Huang, Shan Ping" uniqKey="Huang S" first="Shan-Ping" last="Huang">Shan-Ping Huang</name><name sortKey="Wei, Yen Chiao" sort="Wei, Yen Chiao" uniqKey="Wei Y" first="Yen-Chiao" last="Wei">Yen-Chiao Wei</name><name sortKey="Wu, Shih Lu" sort="Wu, Shih Lu" uniqKey="Wu S" first="Shih-Lu" last="Wu">Shih-Lu Wu</name></noCountry><country name="Taïwan"><noRegion><name sortKey="Ho, Tin Yun" sort="Ho, Tin Yun" uniqKey="Ho T" first="Tin-Yun" last="Ho">Tin-Yun Ho</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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