EGF-receptor specificity for phosphotyrosine-primed substrates provides signal integration with Src

Nat Struct Mol Biol. 2015 Dec;22(12):983-90. doi: 10.1038/nsmb.3117. Epub 2015 Nov 9.


Aberrant activation of the EGF receptor (EGFR) contributes to many human cancers by activating the Ras-MAPK pathway and other pathways. EGFR signaling is augmented by Src-family kinases, but the mechanism is poorly understood. Here, we show that human EGFR preferentially phosphorylates peptide substrates that are primed by a prior phosphorylation. Using peptides based on the sequence of the adaptor protein Shc1, we show that Src mediates the priming phosphorylation, thus promoting subsequent phosphorylation by EGFR. Importantly, the doubly phosphorylated Shc1 peptide binds more tightly than singly phosphorylated peptide to the Ras activator Grb2; this binding is a key step in activating the Ras-MAPK pathway. Finally, a crystal structure of EGFR in complex with a primed Shc1 peptide reveals the structural basis for EGFR substrate specificity. These results provide a molecular explanation for the integration of Src and EGFR signaling with downstream effectors such as Ras.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Crystallography, X-Ray
  • ErbB Receptors / chemistry
  • ErbB Receptors / drug effects*
  • ErbB Receptors / metabolism*
  • GRB2 Adaptor Protein / metabolism
  • Humans
  • Peptides / metabolism*
  • Phosphorylation
  • Phosphotyrosine / metabolism*
  • Protein Binding
  • Protein Conformation
  • Protein Processing, Post-Translational
  • Sensitivity and Specificity
  • Shc Signaling Adaptor Proteins / chemistry
  • Shc Signaling Adaptor Proteins / metabolism*
  • Signal Transduction
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Substrate Specificity


  • GRB2 Adaptor Protein
  • GRB2 protein, human
  • Peptides
  • SHC1 protein, human
  • Shc Signaling Adaptor Proteins
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Phosphotyrosine
  • EGFR protein, human
  • ErbB Receptors

Associated data

  • PDB/5CZH
  • PDB/5CZI