Spike protein, S, of human coronavirus HKU1: role in viral life cycle and application in antibody detection.
Identifieur interne : 003013 ( Main/Exploration ); précédent : 003012; suivant : 003014Spike protein, S, of human coronavirus HKU1: role in viral life cycle and application in antibody detection.
Auteurs : Che-Man Chan [Hong Kong] ; Patrick C Y. Woo ; Susanna K P. Lau ; Herman Tse ; Hong-Lin Chen ; Feng Li ; Bo-Jian Zheng ; Ling Chen ; Jian-Dong Huang ; Kwok-Yung YuenSource :
- Experimental biology and medicine (Maywood, N.J.) [ 1535-3702 ] ; 2008.
Descripteurs français
- KwdFr :
- Animaux, Anticorps antiviraux (génétique), Anticorps antiviraux (immunologie), Anticorps antiviraux (métabolisme), Colorants fluorescents (métabolisme), Coronavirus (), Coronavirus (génétique), Coronavirus (immunologie), Coronavirus (métabolisme), Cricetinae, Fluorescéine-5-isothiocyanate (métabolisme), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (immunologie), Glycoprotéines membranaires (métabolisme), Glycosylation, Humains, Immunisation, Lignée cellulaire, Phénomènes physiologiques (génétique), Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (immunologie), Protéines de l'enveloppe virale (métabolisme), Protéines recombinantes (immunologie), Protéines recombinantes (métabolisme), Rein (cytologie), Souris, Souris de lignée BALB C, Technique d'immunofluorescence indirecte, Transfection.
- MESH :
- cytologie : Rein.
- génétique : Anticorps antiviraux, Coronavirus, Glycoprotéines membranaires, Phénomènes physiologiques, Protéines de l'enveloppe virale.
- immunologie : Anticorps antiviraux, Coronavirus, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines recombinantes.
- métabolisme : Anticorps antiviraux, Colorants fluorescents, Coronavirus, Fluorescéine-5-isothiocyanate, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines recombinantes.
- Animaux, Coronavirus, Cricetinae, Glycoprotéine de spicule des coronavirus, Glycosylation, Humains, Immunisation, Lignée cellulaire, Souris, Souris de lignée BALB C, Technique d'immunofluorescence indirecte, Transfection.
English descriptors
- KwdEn :
- Animals, Antibodies, Viral (genetics), Antibodies, Viral (immunology), Antibodies, Viral (metabolism), Cell Line, Coronavirus (classification), Coronavirus (genetics), Coronavirus (immunology), Coronavirus (metabolism), Cricetinae, Fluorescein-5-isothiocyanate (metabolism), Fluorescent Antibody Technique, Indirect, Fluorescent Dyes (metabolism), Glycosylation, Humans, Immunization, Kidney (cytology), Membrane Glycoproteins (genetics), Membrane Glycoproteins (immunology), Membrane Glycoproteins (metabolism), Mice, Mice, Inbred BALB C, Physiological Phenomena (genetics), Recombinant Proteins (immunology), Recombinant Proteins (metabolism), Spike Glycoprotein, Coronavirus, Transfection, Viral Envelope Proteins (genetics), Viral Envelope Proteins (immunology), Viral Envelope Proteins (metabolism).
- MESH :
- chemical , genetics : Antibodies, Viral, Membrane Glycoproteins, Viral Envelope Proteins.
- chemical , immunology : Antibodies, Viral, Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , metabolism : Antibodies, Viral, Fluorescein-5-isothiocyanate, Fluorescent Dyes, Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- classification : Coronavirus.
- cytology : Kidney.
- genetics : Coronavirus, Physiological Phenomena.
- immunology : Coronavirus.
- metabolism : Coronavirus.
- Animals, Cell Line, Cricetinae, Fluorescent Antibody Technique, Indirect, Glycosylation, Humans, Immunization, Mice, Mice, Inbred BALB C, Spike Glycoprotein, Coronavirus, Transfection.
Abstract
We recently described the discovery, genome, clinical features, genotypes and evolution of a novel and global human respiratory virus named human coronavirus HKU1 (HCoV-HKU1) which is not yet culturable. We expressed a C-terminal FLAG-tagged CoV-HKU1 spike (S) protein by the Semliki Forest Virus (SFV) system and investigated its maturation profile. Pulse chase labeling revealed that S-FLAG was expressed as high-mannose N-glycans of monomers and trimers. It was predominantly cleaved into subdomains S1 and S2 during maturation. S1 was secreted into the medium. Immunofluorescence analysis visualized S along the secretory pathway from endoplasmic reticulum to plasma membrane. Cleavage of S and release of HCoV-HKU1 S pseudotyped virus were inhibited by furin or furin-like enzyme inhibitors. The cell-based expressed full-length S-FLAG could be recognized by the convalescent serum obtained from a patient with HCoV-HKU1 pneumonia. The data suggest that the native form of HCoV-HKU1 spike expressed in our system can be used in developing serological diagnostic assay and in understanding the role of S in the viral life cycle.
DOI: 10.3181/0806-RM-197
PubMed: 18849544
Affiliations:
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Le document en format XML
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Woo</name></author><author><name sortKey="Lau, Susanna K P" sort="Lau, Susanna K P" uniqKey="Lau S" first="Susanna K P" last="Lau">Susanna K P. Lau</name></author><author><name sortKey="Tse, Herman" sort="Tse, Herman" uniqKey="Tse H" first="Herman" last="Tse">Herman Tse</name></author><author><name sortKey="Chen, Hong Lin" sort="Chen, Hong Lin" uniqKey="Chen H" first="Hong-Lin" last="Chen">Hong-Lin Chen</name></author><author><name sortKey="Li, Feng" sort="Li, Feng" uniqKey="Li F" first="Feng" last="Li">Feng Li</name></author><author><name sortKey="Zheng, Bo Jian" sort="Zheng, Bo Jian" uniqKey="Zheng B" first="Bo-Jian" last="Zheng">Bo-Jian Zheng</name></author><author><name sortKey="Chen, Ling" sort="Chen, Ling" uniqKey="Chen L" first="Ling" last="Chen">Ling Chen</name></author><author><name sortKey="Huang, Jian Dong" sort="Huang, Jian Dong" uniqKey="Huang J" first="Jian-Dong" last="Huang">Jian-Dong Huang</name></author><author><name sortKey="Yuen, Kwok Yung" sort="Yuen, Kwok Yung" uniqKey="Yuen K" first="Kwok-Yung" last="Yuen">Kwok-Yung Yuen</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2008">2008</date><idno type="RBID">pubmed:18849544</idno><idno type="pmid">18849544</idno><idno type="doi">10.3181/0806-RM-197</idno><idno type="wicri:Area/PubMed/Corpus">001A53</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001A53</idno><idno type="wicri:Area/PubMed/Curation">001A53</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001A53</idno><idno type="wicri:Area/PubMed/Checkpoint">001A14</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001A14</idno><idno type="wicri:Area/Ncbi/Merge">001D65</idno><idno type="wicri:Area/Ncbi/Curation">001D65</idno><idno type="wicri:Area/Ncbi/Checkpoint">001D65</idno><idno type="wicri:doubleKey">1535-3702:2008:Chan C:spike:protein:s</idno><idno type="wicri:Area/Main/Merge">003083</idno><idno type="wicri:Area/Main/Curation">003013</idno><idno type="wicri:Area/Main/Exploration">003013</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Spike protein, S, of human coronavirus HKU1: role in viral life cycle and application in antibody detection.</title><author><name sortKey="Chan, Che Man" sort="Chan, Che Man" uniqKey="Chan C" first="Che-Man" last="Chan">Che-Man Chan</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, The University of Hong Kong, Hong Kong Special Administration Region, Hong Kong.</nlm:affiliation><country xml:lang="fr">Hong Kong</country><wicri:regionArea>Department of Microbiology, The University of Hong Kong, Hong Kong Special Administration Region</wicri:regionArea><wicri:noRegion>Hong Kong Special Administration Region</wicri:noRegion></affiliation></author><author><name sortKey="Woo, Patrick C Y" sort="Woo, Patrick C Y" uniqKey="Woo P" first="Patrick C Y" last="Woo">Patrick C Y. Woo</name></author><author><name sortKey="Lau, Susanna K P" sort="Lau, Susanna K P" uniqKey="Lau S" first="Susanna K P" last="Lau">Susanna K P. Lau</name></author><author><name sortKey="Tse, Herman" sort="Tse, Herman" uniqKey="Tse H" first="Herman" last="Tse">Herman Tse</name></author><author><name sortKey="Chen, Hong Lin" sort="Chen, Hong Lin" uniqKey="Chen H" first="Hong-Lin" last="Chen">Hong-Lin Chen</name></author><author><name sortKey="Li, Feng" sort="Li, Feng" uniqKey="Li F" first="Feng" last="Li">Feng Li</name></author><author><name sortKey="Zheng, Bo Jian" sort="Zheng, Bo Jian" uniqKey="Zheng B" first="Bo-Jian" last="Zheng">Bo-Jian Zheng</name></author><author><name sortKey="Chen, Ling" sort="Chen, Ling" uniqKey="Chen L" first="Ling" last="Chen">Ling Chen</name></author><author><name sortKey="Huang, Jian Dong" sort="Huang, Jian Dong" uniqKey="Huang J" first="Jian-Dong" last="Huang">Jian-Dong Huang</name></author><author><name sortKey="Yuen, Kwok Yung" sort="Yuen, Kwok Yung" uniqKey="Yuen K" first="Kwok-Yung" last="Yuen">Kwok-Yung Yuen</name></author></analytic><series><title level="j">Experimental biology and medicine (Maywood, N.J.)</title><idno type="ISSN">1535-3702</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Viral (genetics)</term><term>Antibodies, Viral (immunology)</term><term>Antibodies, Viral (metabolism)</term><term>Cell Line</term><term>Coronavirus (classification)</term><term>Coronavirus (genetics)</term><term>Coronavirus (immunology)</term><term>Coronavirus (metabolism)</term><term>Cricetinae</term><term>Fluorescein-5-isothiocyanate (metabolism)</term><term>Fluorescent Antibody Technique, Indirect</term><term>Fluorescent Dyes (metabolism)</term><term>Glycosylation</term><term>Humans</term><term>Immunization</term><term>Kidney (cytology)</term><term>Membrane Glycoproteins (genetics)</term><term>Membrane Glycoproteins (immunology)</term><term>Membrane Glycoproteins (metabolism)</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Physiological Phenomena (genetics)</term><term>Recombinant Proteins (immunology)</term><term>Recombinant Proteins (metabolism)</term><term>Spike Glycoprotein, Coronavirus</term><term>Transfection</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>Animaux</term><term>Anticorps antiviraux (génétique)</term><term>Anticorps antiviraux (immunologie)</term><term>Anticorps antiviraux (métabolisme)</term><term>Colorants fluorescents (métabolisme)</term><term>Coronavirus ()</term><term>Coronavirus (génétique)</term><term>Coronavirus (immunologie)</term><term>Coronavirus (métabolisme)</term><term>Cricetinae</term><term>Fluorescéine-5-isothiocyanate (métabolisme)</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (génétique)</term><term>Glycoprotéines membranaires (immunologie)</term><term>Glycoprotéines membranaires (métabolisme)</term><term>Glycosylation</term><term>Humains</term><term>Immunisation</term><term>Lignée cellulaire</term><term>Phénomènes physiologiques (génétique)</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 (immunologie)</term><term>Protéines recombinantes (métabolisme)</term><term>Rein (cytologie)</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Technique d'immunofluorescence indirecte</term><term>Transfection</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antibodies, Viral</term><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Viral</term><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>Antibodies, Viral</term><term>Fluorescein-5-isothiocyanate</term><term>Fluorescent Dyes</term><term>Membrane Glycoproteins</term><term>Recombinant Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="classification" xml:lang="en"><term>Coronavirus</term></keywords><keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Rein</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Kidney</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Coronavirus</term><term>Physiological Phenomena</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Anticorps antiviraux</term><term>Coronavirus</term><term>Glycoprotéines membranaires</term><term>Phénomènes physiologiques</term><term>Protéines de l'enveloppe virale</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiviraux</term><term>Coronavirus</term><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Coronavirus</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Coronavirus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Anticorps antiviraux</term><term>Colorants fluorescents</term><term>Coronavirus</term><term>Fluorescéine-5-isothiocyanate</term><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cell Line</term><term>Cricetinae</term><term>Fluorescent Antibody Technique, Indirect</term><term>Glycosylation</term><term>Humans</term><term>Immunization</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Spike Glycoprotein, Coronavirus</term><term>Transfection</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Coronavirus</term><term>Cricetinae</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycosylation</term><term>Humains</term><term>Immunisation</term><term>Lignée cellulaire</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Technique d'immunofluorescence indirecte</term><term>Transfection</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">We recently described the discovery, genome, clinical features, genotypes and evolution of a novel and global human respiratory virus named human coronavirus HKU1 (HCoV-HKU1) which is not yet culturable. We expressed a C-terminal FLAG-tagged CoV-HKU1 spike (S) protein by the Semliki Forest Virus (SFV) system and investigated its maturation profile. Pulse chase labeling revealed that S-FLAG was expressed as high-mannose N-glycans of monomers and trimers. It was predominantly cleaved into subdomains S1 and S2 during maturation. S1 was secreted into the medium. Immunofluorescence analysis visualized S along the secretory pathway from endoplasmic reticulum to plasma membrane. Cleavage of S and release of HCoV-HKU1 S pseudotyped virus were inhibited by furin or furin-like enzyme inhibitors. The cell-based expressed full-length S-FLAG could be recognized by the convalescent serum obtained from a patient with HCoV-HKU1 pneumonia. The data suggest that the native form of HCoV-HKU1 spike expressed in our system can be used in developing serological diagnostic assay and in understanding the role of S in the viral life cycle.</div></front></TEI><affiliations><list><country><li>Hong Kong</li></country></list><tree><noCountry><name sortKey="Chen, Hong Lin" sort="Chen, Hong Lin" uniqKey="Chen H" first="Hong-Lin" last="Chen">Hong-Lin Chen</name><name sortKey="Chen, Ling" sort="Chen, Ling" uniqKey="Chen L" first="Ling" last="Chen">Ling Chen</name><name sortKey="Huang, Jian Dong" sort="Huang, Jian Dong" uniqKey="Huang J" first="Jian-Dong" last="Huang">Jian-Dong Huang</name><name sortKey="Lau, Susanna K P" sort="Lau, Susanna K P" uniqKey="Lau S" first="Susanna K P" last="Lau">Susanna K P. Lau</name><name sortKey="Li, Feng" sort="Li, Feng" uniqKey="Li F" first="Feng" last="Li">Feng Li</name><name sortKey="Tse, Herman" sort="Tse, Herman" uniqKey="Tse H" first="Herman" last="Tse">Herman Tse</name><name sortKey="Woo, Patrick C Y" sort="Woo, Patrick C Y" uniqKey="Woo P" first="Patrick C Y" last="Woo">Patrick C Y. Woo</name><name sortKey="Yuen, Kwok Yung" sort="Yuen, Kwok Yung" uniqKey="Yuen K" first="Kwok-Yung" last="Yuen">Kwok-Yung Yuen</name><name sortKey="Zheng, Bo Jian" sort="Zheng, Bo Jian" uniqKey="Zheng B" first="Bo-Jian" last="Zheng">Bo-Jian Zheng</name></noCountry><country name="Hong Kong"><noRegion><name sortKey="Chan, Che Man" sort="Chan, Che Man" uniqKey="Chan C" first="Che-Man" last="Chan">Che-Man Chan</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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