Time-resolved proximity labeling of protein networks associated with ligand-activated EGFR

Cell Rep. 2022 Jun 14;39(11):110950. doi: 10.1016/j.celrep.2022.110950.

Abstract

Ligand binding to the EGF receptor (EGFR) triggers multiple signal-transduction processes and promotes endocytosis of the receptor. The mechanisms of EGFR endocytosis and its cross-talk with signaling are poorly understood. Here, we combine peroxidase-catalyzed proximity labeling, isobaric peptide tagging, and quantitative mass spectrometry to define the dynamics of the proximity proteome of ligand-activated EGFR. Using this approach, we identify a network of signaling proteins, which remain associated with the receptor during its internalization and trafficking through the endosomal system. We show that Trk-fused gene (TFG), a protein known to function at the endoplasmic reticulum exit sites, is enriched in the proximity proteome of EGFR in early/sorting endosomes and localized in these endosomes and demonstrate that TFG regulates endosomal sorting of EGFR. This study provides a comprehensive resource of time-dependent nanoscale environment of EGFR, thus opening avenues to discovering new regulatory mechanisms of signaling and intracellular trafficking of receptor tyrosine kinases.

Keywords: CP: Molecular biology; EGF receptor; endocytosis; proximity labeling; signaling.

Publication types

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

MeSH terms

  • Endocytosis / physiology
  • Endosomes / metabolism
  • Epidermal Growth Factor / metabolism
  • ErbB Receptors* / metabolism
  • Ligands
  • Protein Transport
  • Proteome* / metabolism

Substances

  • Ligands
  • Proteome
  • Epidermal Growth Factor
  • ErbB Receptors