Phospho-tyrosine dependent protein-protein interaction network

Mol Syst Biol. 2015 Mar 26;11(3):794. doi: 10.15252/msb.20145968.


Post-translational protein modifications, such as tyrosine phosphorylation, regulate protein-protein interactions (PPIs) critical for signal processing and cellular phenotypes. We extended an established yeast two-hybrid system employing human protein kinases for the analyses of phospho-tyrosine (pY)-dependent PPIs in a direct experimental, large-scale approach. We identified 292 mostly novel pY-dependent PPIs which showed high specificity with respect to kinases and interacting proteins and validated a large fraction in co-immunoprecipitation experiments from mammalian cells. About one-sixth of the interactions are mediated by known linear sequence binding motifs while the majority of pY-PPIs are mediated by other linear epitopes or governed by alternative recognition modes. Network analysis revealed that pY-mediated recognition events are tied to a highly connected protein module dedicated to signaling and cell growth pathways related to cancer. Using binding assays, protein complementation and phenotypic readouts to characterize the pY-dependent interactions of TSPAN2 (tetraspanin 2) and GRB2 or PIK3R3 (p55γ), we exemplarily provide evidence that the two pY-dependent PPIs dictate cellular cancer phenotypes.

Keywords: cancer signaling; network biology; post‐translational protein modification; yeast two‐hybrid.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Humans
  • Phosphoproteins / metabolism*
  • Phosphorylation
  • Protein Binding
  • Protein Interaction Maps*
  • Protein Kinases / metabolism
  • Protein Processing, Post-Translational
  • Two-Hybrid System Techniques
  • Tyrosine / metabolism*


  • Phosphoproteins
  • Tyrosine
  • Protein Kinases