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Biochemical Characterization of Middle East Respiratory Syndrome Coronavirus Spike Protein Proteolytic Processing.
Whittaker, Gary R; Millet, Jean K.
  • Whittaker GR; Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
  • Millet JK; Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA. jean.millet@inra.fr.
Methods Mol Biol ; 2099: 21-37, 2020.
Article in English | MEDLINE | ID: covidwho-1292545
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Coronavirus Infections / Spike Glycoprotein, Coronavirus / Middle East Respiratory Syndrome Coronavirus Limits: Humans Language: English Journal: Methods Mol Biol Journal subject: Molecular Biology Year: 2020 Document Type: Article Affiliation country: 978-1-0716-0211-9_3

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Coronavirus Infections / Spike Glycoprotein, Coronavirus / Middle East Respiratory Syndrome Coronavirus Limits: Humans Language: English Journal: Methods Mol Biol Journal subject: Molecular Biology Year: 2020 Document Type: Article Affiliation country: 978-1-0716-0211-9_3