Association of pp60src with triton X-100-insoluble residue in human blood platelets requires platelet aggregation and actin polymerization

Atsushi Oda, Brian Druker, Marianne Smith, Edwin W. Salzman

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Protein-tyrosine phosphorylation during platelet activation is inhibited under conditions that inhibit platelet binding of fibrinogen and aggregation. We suggested that pp60src, a major platelet tyrosine kinase, or its protein substrates might become associated with the cytoskeleton upon platelet stimulation, and that this might be related to aggregation. By Western blotting with an anti-Src monoclonal antibody, we found time-dependent association of pp60src with the cytoskeleton (10,000 x g Triton X-100-insoluble matrix) but not the "membrane" cytoskeleton (100,000 x g Triton X-100-insoluble matrix) in platelets activated by U46619 (PGH2 analog). Cytoskeletal association and platelet aggregation were inhibited by the peptide Arg-Gly-Asp-Ser (RGDS) (but not by Arg-Gly-Glu-Ser (RGES)), by 10E5 antibody against glycoprotein (Gp) IIb/IIIa, and by EGTA. U46619-induced association of pp60src with cytoskeleton but not secretion or aggregation was inhibited by cytochalasin D (2 MM). Both cytochalasin D and RGDS inhibited "slow" tyrosine phosphorylation of platelet proteins. Association of pp60src with cytoskeleton induced by U46619 or ADP was not blocked by aspirin. Aspirin blocked epinephrine-induced association of pp60src with the cytoskeleton during a second phase of aggregation when an initial phase had occurred without shape change or secretion. Association of GpIIb/IIIa with the cytoskeleton also accompanied platelet aggregation, shape change, and actin polymerization; this was shown with anti-GpIIb and anti-GpIIIa antibodies. Association of pp60src and GpIIb/IIIa with the cytoskeleton and slow tyrosine phosphorylation are related phenomena.

Original languageEnglish (US)
Pages (from-to)20075-20081
Number of pages7
JournalJournal of Biological Chemistry
Volume267
Issue number28
StatePublished - Oct 5 1992
Externally publishedYes

Fingerprint

Octoxynol
Platelets
Cytoskeleton
Platelet Aggregation
Polymerization
Actins
Blood
Blood Platelets
Agglomeration
15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid
Phosphorylation
Association reactions
Tyrosine
Cytochalasin D
RGES peptide
Aspirin
arginyl-glycyl-aspartyl-serine
Prostaglandin H2
Platelet Glycoprotein GPIIb-IIIa Complex
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Association of pp60src with triton X-100-insoluble residue in human blood platelets requires platelet aggregation and actin polymerization. / Oda, Atsushi; Druker, Brian; Smith, Marianne; Salzman, Edwin W.

In: Journal of Biological Chemistry, Vol. 267, No. 28, 05.10.1992, p. 20075-20081.

Research output: Contribution to journalArticle

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