Direct demonstration of involvement of the adaptor protein ShcA in the regulation of Ca 2+-induced platelet aggregation

Tomohito Higashi, Akira Yoshioka, Ryutaro Shirakawa, Arata Tabuchi, Hiroaki Nishioka, Toru Kita, Hisanori Horiuchi

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Platelet aggregation is mediated by conformational change of integrin α IIbβ 3. Tyrosine-phosphorylation of cytoplasmic domain of β 3 upon platelet activation has been demonstrated to play a critical role in this process. Recently, the adaptor protein ShcA has been shown to bind to the tyrosine-phosphorylated β 3, while it remains open whether ShcA plays any role in platelet aggregation. Here, we show that ShcA bound to tyrosine-phosphorylated β 3-tail peptide through its phosphotyrosine-binding domain in vitro. Then, we examined the involvement of ShcA in platelet aggregation by a previously established in vitro assay using platelets permeabilized with streptolysin-O, where exogenous addition of platelet cytosol is required for reconstitution of the Ca 2+-induced aggregation. When ShcA was specifically depleted with anti-ShcA antibody from the cytosol, this ShcA-depleted cytosol lost the aggregation-supporting activity, which was rescued by addition of purified recombinant ShcA. Thus, ShcA is essential for the Ca 2+-induced platelet aggregation.

Original languageEnglish
Pages (from-to)700-704
Number of pages5
JournalBiochemical and biophysical research communications
Volume322
Issue number2
DOIs
Publication statusPublished - 2004 Sep 17
Externally publishedYes

Keywords

  • Aggregation
  • Integrin
  • Phosphotyrosine-binding domain
  • Platelet
  • ShcA
  • Streptolysin-O

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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