Glycoprotein IIb-IIIa-dependent aggregation by glycoprotein ibα is reinforced by a Src family kinase inhibitor (PP1)-sensitive signalling pathway

Stuart J. Marshall, Naoki Asazuma, Denise Best, Peter Wonerow, Gary Salmon, Robert K. Andrews, Steve P. Watson

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It has been proposed that the receptor for von Willebrand factor (vWF), glycoprotein (GP)Ib-IX-V, signals through the same pathway as the collagen receptor, GPVI, namely via Src kinases, the Fc receptor (FcR) γ-chain and Syk, leading to tyrosine phosphorylation of phospholipase Cγ2 (PLCγ2). The aim of the present study was to assess the functional significance of this pathway in platelet activation by GPIb-IX-V. In washed platelets, vWF/ristocetin and vWF/botrocetin stimulate weak tyrosine phosphorylation of the FcR γ-chain, Syk and PLCγ2, but not the adaptor LAT (linker for activation of T-cells), which is localized to glycolipid-enriched membrane domains. Increases in tyrosine phosphorylation were blocked by the Src family kinase inhibitor, 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo-D-3,4-pyrimidine (PPI). Under the same conditions, neither stimulus induced activation of PLCγ2 nor functional responses, such as Ca2+ elevation, secretion or GPIIb-IIIa-dependent aggregation. In contrast, in platelet-rich plasma (PRP), threshold concentrations of ristocetin or asialo-vWF stimulated GPIb-dependent biphasic aggregation, in which the second phase was blocked by PP1. Importantly, a significant component of the initial phase and the complete second phase of aggregation was blocked by GPIIb-IIIa receptor antagonists in PRP. Higher concentrations of ristocetin stimulated GPIIb-IIIa-independent agglutination in PRP. These results demonstrate that GPIb-IX-V initiates activation of GPIIb-IIIa in PRP through an undefined pathway that is reinforced by a PP1-sensitive pathway. In contrast, activation of GPIbα in washed platelets does not promote functional responses.

Original languageEnglish
Pages (from-to)297-305
Number of pages9
JournalBiochemical Journal
Issue number2
Publication statusPublished - 15 Jan 2002
Externally publishedYes


  • Src kinases
  • Tyrosine phosphorylation
  • Von Willebrand factor

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