Identification of protein receptors for coronaviruses by mass spectrometry

Methods Mol Biol. 2015;1282:165-82. doi: 10.1007/978-1-4939-2438-7_15.

Abstract

As obligate intracellular parasites, viruses need to cross the plasma membrane and deliver their genome inside the cell. This step is initiated by the recognition of receptors present on the host cell surface. Receptors can be major determinants of tropism, host range, and pathogenesis. Identifying virus receptors can give clues to these aspects and can lead to the design of intervention strategies. Interfering with receptor recognition is an attractive antiviral therapy, since it occurs before the viral genome has reached the relative safe haven within the cell. This chapter describes the use of an immunoprecipitation approach with Fc-tagged viral spike proteins followed by mass spectrometry to identify and characterize the receptor for the Middle East respiratory syndrome coronavirus. This technique can be adapted to identify other viral receptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • COS Cells
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Coronavirus / physiology*
  • DNA Primers / genetics
  • Dipeptidyl Peptidase 4 / genetics
  • Dipeptidyl Peptidase 4 / isolation & purification
  • Dipeptidyl Peptidase 4 / metabolism
  • Flow Cytometry
  • HEK293 Cells
  • Host-Pathogen Interactions
  • Humans
  • Immunoglobulin G / biosynthesis
  • Immunoglobulin G / genetics
  • Immunoprecipitation
  • Receptors, Virus / genetics
  • Receptors, Virus / isolation & purification
  • Receptors, Virus / metabolism*
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Spike Glycoprotein, Coronavirus / biosynthesis
  • Spike Glycoprotein, Coronavirus / genetics
  • Tandem Mass Spectrometry
  • Virus Internalization

Substances

  • DNA Primers
  • Immunoglobulin G
  • Receptors, Virus
  • Recombinant Fusion Proteins
  • Spike Glycoprotein, Coronavirus
  • DPP4 protein, human
  • Dipeptidyl Peptidase 4