Tyrosine residues 239 and 240 of Shc are phosphatidylinositol 4,5-bisphosphate-dependent phosphorylation sites by c-Src

Biochem Biophys Res Commun. 1997 Nov 17;240(2):399-404. doi: 10.1006/bbrc.1997.7667.


In the previous study (Sato K.-I. et al. (1997) FEBS Lett. 410, 136-140), we showed that the phosphorylation of Shc protein by c-Src is dependent on the binding of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) to the PTB domain of Shc. In this study, we demonstrate that, in contrast to c-Src, v-Src and epidermal growth factor (EGF) receptor can phosphorylate Shc in a PtdIns(4,5)P2-independent manner and at different phosphorylation sites. To determine the phosphorylation sites in Shc, we used mutant Shc proteins in which tyrosine residues (Y) 317 and/or 239 and 240 were replaced by phenylalanine residues (F). We found that Y317F Shc but not Y239/240F or Y239/240/317F Shc was phosphorylated by c-Src. The reaction was PtdIns(4,5)P2-dependent and inhibited by the addition of PTB domain of Shc. On the other hand, v-Src and EGF receptor were able to phosphorylate both Y317F and Y239/240F but not Y239/240/317F Shc in a PtdIns(4,5)P2-independent manner. These results highlight the difference between c-Src and v-Src or EGF receptor and suggest that c-Src can phosphorylate predominantly on Tyr239/240 of Shc only when Shc PTB domain is bound to PtdIns(4,5)P2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Adaptor Proteins, Vesicular Transport*
  • Amino Acid Substitution
  • Animals
  • Brain / metabolism
  • Carcinoma, Squamous Cell
  • Cattle
  • Cell Line, Transformed
  • ErbB Receptors / metabolism
  • Glutathione Transferase
  • Humans
  • Mice
  • Mutagenesis, Site-Directed
  • Oncogene Protein pp60(v-src) / metabolism
  • Phosphatidylinositol 4,5-Diphosphate / metabolism*
  • Phosphatidylinositol 4,5-Diphosphate / pharmacology*
  • Protein Kinases / metabolism
  • Proteins / chemistry*
  • Proteins / metabolism*
  • Proto-Oncogene Proteins pp60(c-src) / metabolism*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Shc Signaling Adaptor Proteins
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Tumor Cells, Cultured
  • Tyrosine*


  • Adaptor Proteins, Signal Transducing
  • Adaptor Proteins, Vesicular Transport
  • Phosphatidylinositol 4,5-Diphosphate
  • Proteins
  • Recombinant Fusion Proteins
  • SHC1 protein, human
  • Shc Signaling Adaptor Proteins
  • Shc1 protein, mouse
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Tyrosine
  • Glutathione Transferase
  • Protein Kinases
  • ErbB Receptors
  • Oncogene Protein pp60(v-src)
  • Proto-Oncogene Proteins pp60(c-src)