Endocytosis of the receptor-binding domain of SARS-CoV spike protein together with virus receptor ACE2.
Identifieur interne : 003118 ( Main/Exploration ); précédent : 003117; suivant : 003119Endocytosis of the receptor-binding domain of SARS-CoV spike protein together with virus receptor ACE2.
Auteurs : Shunxin Wang [République populaire de Chine] ; Feng Guo ; Kangtai Liu ; Hongliang Wang ; Shuan Rao ; Peng Yang ; Chengyu JiangSource :
- Virus research [ 0168-1702 ] ; 2008.
Descripteurs français
- KwdFr :
- Animaux, Cytométrie en flux, Endocytose, Fragments Fc des immunoglobulines (génétique), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (métabolisme), Humains, Liaison aux protéines, Lignée cellulaire, Peptidyl-Dipeptidase A (métabolisme), Protéines de fusion recombinantes (génétique), Protéines de fusion recombinantes (métabolisme), Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (métabolisme), Récepteurs viraux (métabolisme), Virus du SRAS (physiologie).
- MESH :
- génétique : Fragments Fc des immunoglobulines, Glycoprotéines membranaires, Protéines de fusion recombinantes, Protéines de l'enveloppe virale.
- métabolisme : Glycoprotéines membranaires, Peptidyl-Dipeptidase A, Protéines de fusion recombinantes, Protéines de l'enveloppe virale, Récepteurs viraux.
- physiologie : Virus du SRAS.
- Animaux, Cytométrie en flux, Endocytose, Glycoprotéine de spicule des coronavirus, Humains, Liaison aux protéines, Lignée cellulaire.
English descriptors
- KwdEn :
- Animals, Cell Line, Chlorocebus aethiops, Endocytosis, Flow Cytometry, Humans, Immunoglobulin Fc Fragments (genetics), Membrane Glycoproteins (genetics), Membrane Glycoproteins (metabolism), Peptidyl-Dipeptidase A (metabolism), Protein Binding, Receptors, Virus (metabolism), Recombinant Fusion Proteins (genetics), Recombinant Fusion Proteins (metabolism), SARS Virus (physiology), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (genetics), Viral Envelope Proteins (metabolism).
- MESH :
- chemical , genetics : Immunoglobulin Fc Fragments, Membrane Glycoproteins, Recombinant Fusion Proteins, Viral Envelope Proteins.
- chemical , metabolism : Membrane Glycoproteins, Peptidyl-Dipeptidase A, Receptors, Virus, Recombinant Fusion Proteins, Viral Envelope Proteins.
- physiology : SARS Virus.
- Animals, Cell Line, Chlorocebus aethiops, Endocytosis, Flow Cytometry, Humans, Protein Binding, Spike Glycoprotein, Coronavirus.
Abstract
Cell entry of severe acute respiratory syndrome coronavirus (SARS-CoV) is mediated by the viral spike (S) protein. Amino acids 319-510 on the S protein have been mapped as the receptor-binding domain (RBD), which mediates binding to the SARS-CoV receptor angiotensin converting enzyme 2 (ACE2) on SARS-CoV susceptible cells. In this study, we expressed a fusion protein containing the human codon-optimized RBD of the SARS-CoV spike protein linked to the Fc portion of human IgG1 (named RBD-Fc) in HEK293 cells. The RBD-Fc protein was purified by affinity chromatography. The flow cytometry assay showed that the purified RBD-Fc protein could bind to ACE2. We demonstrated that the RBD spike protein alone could be internalized into SARS-CoV susceptible cells together with ACE2. We also showed that the removal of N-glycans from the RBD spike protein did not abolish this phenomenon. Our discoveries may have some implications for the development of the SARS vaccine.
DOI: 10.1016/j.virusres.2008.03.004
PubMed: 18554741
Affiliations:
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Le document en format XML
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spike protein together with virus receptor ACE2.</title><author><name sortKey="Wang, Shunxin" sort="Wang, Shunxin" uniqKey="Wang S" first="Shunxin" last="Wang">Shunxin Wang</name><affiliation wicri:level="1"><nlm:affiliation>National Key Laboratory of Medical Molecular Biology, School of Basic Medicine, Peking Union Medical College, Tsinghua University and Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, Beijing 100005, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Key Laboratory of Medical Molecular Biology, School of Basic Medicine, Peking Union Medical College, Tsinghua University and Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, Beijing 100005</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Guo, Feng" sort="Guo, Feng" uniqKey="Guo F" first="Feng" last="Guo">Feng Guo</name></author><author><name sortKey="Liu, Kangtai" sort="Liu, Kangtai" uniqKey="Liu K" first="Kangtai" last="Liu">Kangtai Liu</name></author><author><name sortKey="Wang, Hongliang" sort="Wang, Hongliang" uniqKey="Wang H" first="Hongliang" last="Wang">Hongliang Wang</name></author><author><name sortKey="Rao, Shuan" sort="Rao, Shuan" uniqKey="Rao S" first="Shuan" last="Rao">Shuan Rao</name></author><author><name sortKey="Yang, Peng" sort="Yang, Peng" uniqKey="Yang P" first="Peng" last="Yang">Peng Yang</name></author><author><name sortKey="Jiang, Chengyu" sort="Jiang, Chengyu" uniqKey="Jiang C" first="Chengyu" last="Jiang">Chengyu Jiang</name></author></analytic><series><title level="j">Virus research</title><idno type="ISSN">0168-1702</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>Cell Line</term><term>Chlorocebus aethiops</term><term>Endocytosis</term><term>Flow Cytometry</term><term>Humans</term><term>Immunoglobulin Fc Fragments (genetics)</term><term>Membrane Glycoproteins (genetics)</term><term>Membrane Glycoproteins (metabolism)</term><term>Peptidyl-Dipeptidase A (metabolism)</term><term>Protein Binding</term><term>Receptors, Virus (metabolism)</term><term>Recombinant Fusion Proteins (genetics)</term><term>Recombinant Fusion Proteins (metabolism)</term><term>SARS Virus (physiology)</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (genetics)</term><term>Viral Envelope Proteins (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Cytométrie en flux</term><term>Endocytose</term><term>Fragments Fc des immunoglobulines (génétique)</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (génétique)</term><term>Glycoprotéines membranaires (métabolisme)</term><term>Humains</term><term>Liaison aux protéines</term><term>Lignée cellulaire</term><term>Peptidyl-Dipeptidase A (métabolisme)</term><term>Protéines de fusion recombinantes (génétique)</term><term>Protéines de fusion recombinantes (métabolisme)</term><term>Protéines de l'enveloppe virale (génétique)</term><term>Protéines de l'enveloppe virale (métabolisme)</term><term>Récepteurs viraux (métabolisme)</term><term>Virus du SRAS (physiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Immunoglobulin Fc Fragments</term><term>Membrane Glycoproteins</term><term>Recombinant Fusion Proteins</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>Recombinant Fusion Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Fragments Fc des immunoglobulines</term><term>Glycoprotéines membranaires</term><term>Protéines de fusion recombinantes</term><term>Protéines de l'enveloppe virale</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Peptidyl-Dipeptidase A</term><term>Protéines de fusion recombinantes</term><term>Protéines de l'enveloppe virale</term><term>Récepteurs viraux</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cell Line</term><term>Chlorocebus aethiops</term><term>Endocytosis</term><term>Flow Cytometry</term><term>Humans</term><term>Protein Binding</term><term>Spike Glycoprotein, Coronavirus</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cytométrie en flux</term><term>Endocytose</term><term>Glycoprotéine de spicule des coronavirus</term><term>Humains</term><term>Liaison aux protéines</term><term>Lignée cellulaire</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Cell entry of severe acute respiratory syndrome coronavirus (SARS-CoV) is mediated by the viral spike (S) protein. Amino acids 319-510 on the S protein have been mapped as the receptor-binding domain (RBD), which mediates binding to the SARS-CoV receptor angiotensin converting enzyme 2 (ACE2) on SARS-CoV susceptible cells. In this study, we expressed a fusion protein containing the human codon-optimized RBD of the SARS-CoV spike protein linked to the Fc portion of human IgG1 (named RBD-Fc) in HEK293 cells. The RBD-Fc protein was purified by affinity chromatography. The flow cytometry assay showed that the purified RBD-Fc protein could bind to ACE2. We demonstrated that the RBD spike protein alone could be internalized into SARS-CoV susceptible cells together with ACE2. We also showed that the removal of N-glycans from the RBD spike protein did not abolish this phenomenon. Our discoveries may have some implications for the development of the SARS vaccine.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Guo, Feng" sort="Guo, Feng" uniqKey="Guo F" first="Feng" last="Guo">Feng Guo</name><name sortKey="Jiang, Chengyu" sort="Jiang, Chengyu" uniqKey="Jiang C" first="Chengyu" last="Jiang">Chengyu Jiang</name><name sortKey="Liu, Kangtai" sort="Liu, Kangtai" uniqKey="Liu K" first="Kangtai" last="Liu">Kangtai Liu</name><name sortKey="Rao, Shuan" sort="Rao, Shuan" uniqKey="Rao S" first="Shuan" last="Rao">Shuan Rao</name><name sortKey="Wang, Hongliang" sort="Wang, Hongliang" uniqKey="Wang H" first="Hongliang" last="Wang">Hongliang Wang</name><name sortKey="Yang, Peng" sort="Yang, Peng" uniqKey="Yang P" first="Peng" last="Yang">Peng Yang</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Wang, Shunxin" sort="Wang, Shunxin" uniqKey="Wang S" first="Shunxin" last="Wang">Shunxin Wang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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