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Biochemical Characterization of Middle East Respiratory Syndrome Coronavirus Spike Protein Proteolytic Processing

Authors
  • Whittaker, Gary R.1
  • Millet, Jean K.1, 2
  • 1 Cornell University,
  • 2 INRA,
Type
Published Article
Journal
MERS Coronavirus
Publication Date
Sep 14, 2019
Volume
2099
Pages
21–37
Identifiers
DOI: 10.1007/978-1-0716-0211-9_3
PMID: 31883085
PMCID: PMC7121948
Source
PubMed Central
Keywords
License
Unknown

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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