Identification of c-erbB-3 binding sites for phosphatidylinositol 3'-kinase and SHC using an EGF receptor/c-erbB-3 chimera

EMBO J. 1994 Jun 15;13(12):2831-41.

Abstract

c-erbB-3 is a member of the type I (EGF receptor-related) family of growth factor receptors for which no ligand has been identified. To facilitate ligand stimulation we have constructed a chimeric receptor which possesses an activatable kinase and promotes the growth of NIH 3T3 fibroblasts. In this study we have shown that SHC and phosphatidylinositol 3'-kinase bind to the activated EGF receptor/c-erbB-3 chimera. Whereas p85 is not phosphorylated to a significant extent, SHC appears to be a major substrate for phosphorylation on tyrosine. In contrast to EGF receptor and c-erbB-2, we were unable to detect binding of activated c-erbB-3 to GRB2. Using synthetic peptides corresponding to each of 13 potential phosphorylation sites on c-erbB-3, we have shown that tyrosine 1309 is responsible for SHC binding. Peptides containing the motif YXXM inhibit p85 association. By comparison with recently reported SHC binding sites on Middle T antigen and Trk we have identified a SHC binding motif, NPXY.

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Amino Acid Sequence
  • Animals
  • Avian Proteins*
  • Base Sequence
  • Cell Adhesion
  • Cells, Cultured
  • Cytoskeletal Proteins / metabolism
  • ErbB Receptors / genetics
  • ErbB Receptors / metabolism*
  • GRB2 Adaptor Protein
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism
  • Molecular Sequence Data
  • Phosphatidylinositol 3-Kinases
  • Phosphopeptides / genetics
  • Phosphoproteins / metabolism
  • Phosphorylation
  • Phosphotransferases (Alcohol Group Acceptor) / metabolism*
  • Protein Binding
  • Proteins / genetics
  • Proteins / metabolism
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Second Messenger Systems*

Substances

  • Adaptor Proteins, Signal Transducing
  • Avian Proteins
  • Cytoskeletal Proteins
  • GRB2 Adaptor Protein
  • Phosphopeptides
  • Phosphoproteins
  • Proteins
  • Proto-Oncogene Proteins
  • Recombinant Fusion Proteins
  • cytoskeletal p85 protein, chicken
  • Glutathione Transferase
  • Phosphatidylinositol 3-Kinases
  • Phosphotransferases (Alcohol Group Acceptor)
  • ErbB Receptors