Thrombin-stimulated immunoprecipitation of phosphatidylinositol 3-kinase from human platelets

C. A. Mitchell, A. B. Jefferson, B. E. Bejeck, J. S. Brugge, T. F. Deuel, P. W. Majerus

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7 Citations (Scopus)

Abstract

Growth factors and transforming proteins that activate tyrosine phosphorylation have been shown to cause an increased labeling of 3-phosphate-containing phosphatidylinositols. Turnover correlates with the formation of a complex between phosphatidylinositol 3-kinase, the activated protein-tyrosine kinase, and other proteins thought to participate in transmembrane signaling. When human platelets are treated with thrombin, labeling of 3-phosphate-containing phosphatidylinositols is stimulated with a time course and concentration dependence consistent with a role for these lipids in platelet activation. We now report that when human platelets are stimulated with thrombin, a complex forms between phosphatidylinositol 3-kinase, a protein-serine/threonine kinase, and an uncharacterized platelet membrane protein. The complex is immunoprecipitated from detergent lysates of thrombin-stimulated platelets by a rabbit antiserum prepared against a peptide from the cytoplasmic domain of the mouse platelet-derived growth factor (PDGF) receptor. The antigen is not the PDGF receptor, since complex formation is not stimulated by PDGF and thrombin-induced complexes are not precipitated by another rabbit antiserum against the same peptide or by monoclonal anti-human PDGF receptor antibodies. Formation of the complex is rapid (within 30 sec) and occurs at thrombin concentrations that stimulate platelet aggregation and secretion (50% of maximal complex formation at 0.03 unit of thrombin per ml). We propose that the complex initiates formation of 3-phosphate-containing phosphatidylinositols that may function in platelet activation.

Original languageEnglish
Pages (from-to)9396-9400
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number23
DOIs
Publication statusPublished - 1990
Externally publishedYes

Keywords

  • c-src gene
  • protein-serine
  • threonine kinase

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