Chemical proteomics tracks virus entry and uncovers NCAM1 as Zika virus receptor

Nat Commun. 2020 Aug 4;11(1):3896. doi: 10.1038/s41467-020-17638-y.

Abstract

The outbreak of Zika virus (ZIKV) in 2016 created worldwide health emergency which demand urgent research efforts on understanding the virus biology and developing therapeutic strategies. Here, we present a time-resolved chemical proteomic strategy to track the early-stage entry of ZIKV into host cells. ZIKV was labeled on its surface with a chemical probe, which carries a photocrosslinker to covalently link virus-interacting proteins in living cells on UV exposure at different time points, and a biotin tag for subsequent enrichment and mass spectrometric identification of the receptor or other host proteins critical for virus internalization. We identified Neural Cell Adhesion Molecule (NCAM1) as a potential ZIKV receptor and further validated it through overexpression, knockout, and inhibition of NCAM1 in Vero cells and human glioblastoma cells U-251 MG. Collectively, the strategy can serve as a universal tool to map virus entry pathways and uncover key interacting proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • CD56 Antigen / genetics
  • CD56 Antigen / metabolism
  • Cell Line, Tumor
  • Chlorocebus aethiops
  • Gene Knockout Techniques
  • Glioblastoma
  • HEK293 Cells
  • Host-Pathogen Interactions / physiology
  • Humans
  • Neural Cell Adhesion Molecules / genetics
  • Neural Cell Adhesion Molecules / metabolism*
  • Proteomics*
  • Receptors, Virus / metabolism*
  • Vero Cells
  • Viral Proteins / metabolism
  • Virus Attachment
  • Virus Internalization*
  • Virus Replication / physiology*
  • Zika Virus / physiology*
  • Zika Virus Infection / virology

Substances

  • CD56 Antigen
  • NCAM1 protein, human
  • Neural Cell Adhesion Molecules
  • Receptors, Virus
  • Viral Proteins