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Thromb Haemost 2004; 91(01): 102-110
DOI: 10.1160/TH03-02-0083
Platelets and Blood Cells
Schattauer GmbH

A role for p38 MAP kinase in platelet activation by von Willebrand Factor

Ilaria Canobbio
1   Center of Excellence on Applied Biology and Department of Biochemistry, University of Pavia, Italy
,
Stefania Reineri
2   Department of Medical Sciences, University “A. Avogadro”, Novara, Italy
,
Fabiola Sinigaglia
2   Department of Medical Sciences, University “A. Avogadro”, Novara, Italy
,
Cesare Balduini
1   Center of Excellence on Applied Biology and Department of Biochemistry, University of Pavia, Italy
,
Mauro Torti
1   Center of Excellence on Applied Biology and Department of Biochemistry, University of Pavia, Italy
› Author AffiliationsFinancial support: This work was supported by grants from Consiglio Nazionale delle Ricerche (CNR, target project Biotechnology), MIUR (PRIN 2001 and PRIN 2002), and Consorzio Interuniversitario Biotecnologie (CIB). This paper is dedicated to the memory of Eraldo Antonini, eminent biochemist, prematurely deceased twenty years ago, on March 19th 1983.
Further Information

Publication History

Received 06 February 2003

Accepted after resubmission 01 October 2003

Publication Date:
30 November 2017 (online)

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Summary

Platelet activation induced by von Willebrand factor (VWF) binding to the membrane GPIb-IX-V receptor involves multiple signal transduction pathways. Among these, recruitment and activation of the FcγRIIA and stimulation of phospholipase A2 represent independent events equally essential to support a complete platelet response. Phospholipase A2 is activated by calcium and by phosphorylation through MAP kinases. In this work, we found that VWF stimulated the rapid and sustained phosphorylation of p38 MAP kinase (p38MAPK). In vitro kinase assay revealed that VWF-stimulated phosphorylation of p38MAPK was associated with increased kinase activity. Binding of VWF to GPIb-IX-V, but not to integrin αIIbβ3, was required to support phosphorylation of p38MAPK. Neither the blockade of the membrane FcγRIIA by a specific monoclonal antibody or the prevention of thromboxane A2 synthesis by cyclooxygenase inhibitors affected VWF-induced p38MAPK activation. However, phosphorylation of p38MAPK was prevented by the tyrosine kinase Syk inhibitor piceatannol. Treatment of platelets with the p38MAPK inhibitor SB203580 totally prevented VWFstimulated platelet aggregation. Moreover, release of arachidonic acid induced by VWF was strongly impaired by inhibition of p38MAPK. We also found thatVWF induced phosphorylation of cytosolic phospholipase A2, and that this process was prevented by the p38MAPK inhibitor SB203580.These results demonstrate that p38MAPK is a key element in the FcγRIIA-independent pathway for VWF-induced platelet activation, and is involved in the stimulation of phospholipase A2 and arachidonic acid release.

Keywords

p38 MAP kinase - von Willebrand Factor - tyrosine phosphorylation - phospholipase A2 - arachidonic acid
 

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