Regulation of postaggregation events induced by protease-activated receptor 1 ligation in human platelets: Evidence of differential signaling pathways

Amanchy S S Ramars, Saikat Mukhopadhyay, Debabrata Dash

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

In a physiological milieu platelets continue to be exposed to agonists long after clot formation. We studied the regulation of postaggregation events consequent on protease-activated receptor (PAR) 1 ligation with either thrombin or the thrombin receptor-activating peptide (TRAP). Stimulation with TRAP (20 μM) but not with thrombin (1 U/ml) for 15 min evoked platelet disaggregation by about 30% and downregulation of high-affinity fibrinogen binding sites on integrin αIIbβ3 to nearly prestimulation levels. Concurrently, only TRAP disorganized the actin-based cytoskeleton, with decrease in the cytoskeletal content of focal contact-associated proteins like integrin αIIbβ3, Src, and focal adhesion kinase (FAK). While protein tyrosine kinases were activated during the initial period of platelet aggregation with either agonist, stimulation of protein tyrosine phosphatases determined the successive phase of reduced phosphotyrosine content. SHP-1, an abundant protein tyrosine phosphatase in the platelets, was tyrosine phosphorylated on challenge of PAR-1 and coprecipitated with two unidentified tyrosine phosphorylated proteins of 140 and 60 kDa; in addition, SHP-1 tyrosine phosphorylation (which is associated with enhanced phosphatase activity) was sustained until 15 min. Activity of calpain was upregulated following incubation with thrombin and not with TRAP. Collectively, these data suggest that signaling pathways elicited by PAR-1 agonists thrombin and TRAP are markedly different, which could have important implications on late platelet responses.

Original languageEnglish (US)
Pages (from-to)253-260
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume398
Issue number2
DOIs
StatePublished - Feb 15 2002

Keywords

  • Calpain
  • Cytoskeleton
  • Integrin αβ
  • Platelet
  • Protease-activated receptor 1
  • Protein tyrosine kinases
  • Protein tyrosine phosphatases
  • Thrombin

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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