Biochemical Characterization of Middle East Respiratory Syndrome Coronavirus Spike Protein Proteolytic Processing.
Identifieur interne : 000315 ( PubMed/Corpus ); précédent : 000314; suivant : 000316Biochemical Characterization of Middle East Respiratory Syndrome Coronavirus Spike Protein Proteolytic Processing.
Auteurs : Gary R. Whittaker ; Jean K. MilletSource :
- Methods in molecular biology (Clifton, N.J.) [ 1940-6029 ] ; 2020.
Abstract
The coronavirus spike envelope glycoprotein is an essential viral component that mediates virus entry events. Biochemical assessment of the spike protein is critical for understanding structure-function relationships and the roles of the protein in the viral life cycle. Coronavirus spike proteins are typically proteolytically processed and activated by host cell enzymes such as trypsin-like proteases, cathepsins, or proprotein-convertases. Analysis of coronavirus spike proteins by western blot allows the visualization and assessment of proteolytic processing by endogenous or exogenous proteases. Here, we present a method based on western blot analysis to investigate spike protein proteolytic cleavage by transient transfection of HEK-293 T cells allowing expression of the spike protein of the highly pathogenic Middle East respiratory syndrome coronavirus in the presence or absence of a cellular trypsin-like transmembrane serine protease, matriptase. Such analysis enables the characterization of cleavage patterns produced by a host protease on a coronavirus spike glycoprotein.
DOI: 10.1007/978-1-0716-0211-9_3
PubMed: 31883085
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<author><name sortKey="Whittaker, Gary R" sort="Whittaker, Gary R" uniqKey="Whittaker G" first="Gary R" last="Whittaker">Gary R. Whittaker</name>
<affiliation><nlm:affiliation>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.</nlm:affiliation>
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<author><name sortKey="Millet, Jean K" sort="Millet, Jean K" uniqKey="Millet J" first="Jean K" last="Millet">Jean K. Millet</name>
<affiliation><nlm:affiliation>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA. jean.millet@inra.fr.</nlm:affiliation>
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<front><div type="abstract" xml:lang="en">The coronavirus spike envelope glycoprotein is an essential viral component that mediates virus entry events. Biochemical assessment of the spike protein is critical for understanding structure-function relationships and the roles of the protein in the viral life cycle. Coronavirus spike proteins are typically proteolytically processed and activated by host cell enzymes such as trypsin-like proteases, cathepsins, or proprotein-convertases. Analysis of coronavirus spike proteins by western blot allows the visualization and assessment of proteolytic processing by endogenous or exogenous proteases. Here, we present a method based on western blot analysis to investigate spike protein proteolytic cleavage by transient transfection of HEK-293 T cells allowing expression of the spike protein of the highly pathogenic Middle East respiratory syndrome coronavirus in the presence or absence of a cellular trypsin-like transmembrane serine protease, matriptase. Such analysis enables the characterization of cleavage patterns produced by a host protease on a coronavirus spike glycoprotein.</div>
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<Abstract><AbstractText>The coronavirus spike envelope glycoprotein is an essential viral component that mediates virus entry events. Biochemical assessment of the spike protein is critical for understanding structure-function relationships and the roles of the protein in the viral life cycle. Coronavirus spike proteins are typically proteolytically processed and activated by host cell enzymes such as trypsin-like proteases, cathepsins, or proprotein-convertases. Analysis of coronavirus spike proteins by western blot allows the visualization and assessment of proteolytic processing by endogenous or exogenous proteases. Here, we present a method based on western blot analysis to investigate spike protein proteolytic cleavage by transient transfection of HEK-293 T cells allowing expression of the spike protein of the highly pathogenic Middle East respiratory syndrome coronavirus in the presence or absence of a cellular trypsin-like transmembrane serine protease, matriptase. Such analysis enables the characterization of cleavage patterns produced by a host protease on a coronavirus spike glycoprotein.</AbstractText>
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<KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="Y">Coronavirus</Keyword>
<Keyword MajorTopicYN="Y">Host cell protease</Keyword>
<Keyword MajorTopicYN="Y">Matriptase</Keyword>
<Keyword MajorTopicYN="Y">Middle East respiratory syndrome (MERS)</Keyword>
<Keyword MajorTopicYN="Y">Proteolytic processing</Keyword>
<Keyword MajorTopicYN="Y">Spike protein</Keyword>
<Keyword MajorTopicYN="Y">Transient transfection</Keyword>
<Keyword MajorTopicYN="Y">Virus entry</Keyword>
<Keyword MajorTopicYN="Y">Western blot</Keyword>
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