Roles of the complex formation of SHPS-1 with SHP-2 in insulin-stimulated mitogen-activated protein kinase activation

J Biol Chem. 1998 Apr 10;273(15):9234-42. doi: 10.1074/jbc.273.15.9234.

Abstract

SHPS-1 is a receptor-like protein that undergoes tyrosine phosphorylation and binds SHP-2, an SH2 domain-containing protein tyrosine phosphatase, in response to insulin and other mitogens. The overexpression of wild-type SHPS-1, but not of a mutant SHPS-1 in which all four tyrosine residues in its cytoplasmic region were mutated to phenylalanine, markedly enhanced insulin-induced activation of mitogen-activated protein kinase in Chinese hamster ovary cells that overexpress the human insulin receptor. Mutation of each tyrosine residue individually revealed that the major sites of tyrosine phosphorylation of SHPS-1 in response to insulin are Tyr449 and Tyr473. In addition, mutation of either Tyr449 or Tyr473 abolished the insulin-induced tyrosine phosphorylation of SHPS-1 and its association with SHP-2. Surface plasmon resonance analysis showed that glutathione S-transferase fusion proteins containing the NH2-terminal or COOH-terminal SH2 domains of SHP-2 bound preferentially to phosphotyrosyl peptides corresponding to the sequences surrounding Tyr449 or Tyr473, respectively, of SHPS-1. Furthermore, phosphotyrosyl peptides containing Tyr449 or Tyr473 were effective substrates for the phosphatase activity of recombinant SHP-2 in vitro. Together, these results suggest that insulin may induce phosphorylation of SHPS-1 at Tyr449 and Tyr473, to which SHP-2 then binds through its NH2-terminal and COOH-terminal SH2 domains, respectively. SHPS-1 may play a crucial role both in the recruitment of SHP-2 from the cytosol to a site near the plasma membrane and in increasing its catalytic activity, thereby positively regulating the RAS-mitogen-activated protein kinase signaling cascade in response to insulin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antigens, Differentiation*
  • Binding Sites
  • CHO Cells
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
  • Cricetinae
  • DNA Primers
  • Enzyme Activation
  • Humans
  • Insulin / pharmacology*
  • Intracellular Signaling Peptides and Proteins
  • Kinetics
  • Membrane Glycoproteins / biosynthesis
  • Membrane Glycoproteins / isolation & purification
  • Membrane Glycoproteins / metabolism*
  • Mutagenesis, Site-Directed
  • Neural Cell Adhesion Molecule L1*
  • Neural Cell Adhesion Molecules / biosynthesis
  • Neural Cell Adhesion Molecules / isolation & purification
  • Neural Cell Adhesion Molecules / metabolism*
  • Peptide Fragments / chemistry
  • Polymerase Chain Reaction
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6
  • Protein Tyrosine Phosphatases / biosynthesis
  • Protein Tyrosine Phosphatases / isolation & purification
  • Protein Tyrosine Phosphatases / metabolism*
  • Receptor, Insulin / biosynthesis
  • Receptor, Insulin / physiology*
  • Receptors, Immunologic*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • SH2 Domain-Containing Protein Tyrosine Phosphatases
  • Transfection
  • src Homology Domains

Substances

  • Antigens, Differentiation
  • DNA Primers
  • Insulin
  • Intracellular Signaling Peptides and Proteins
  • Membrane Glycoproteins
  • Neural Cell Adhesion Molecule L1
  • Neural Cell Adhesion Molecules
  • Peptide Fragments
  • Receptors, Immunologic
  • Recombinant Proteins
  • SIRPA protein, human
  • Receptor, Insulin
  • Calcium-Calmodulin-Dependent Protein Kinases
  • PTPN11 protein, human
  • PTPN6 protein, human
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6
  • Protein Tyrosine Phosphatases
  • SH2 Domain-Containing Protein Tyrosine Phosphatases