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

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 α_IIbβ₃, 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

• References

• 1 Widmann C, Gibson S, Jarpe MB. et al. Mitogen-activated protein kinase: conservation of three-kinase module from yeast to human. Physiol Rev 1999; 79: 143-80.
• 2 Johnson GL, Lapadat R. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 2002; 298: 1911-12.
• 3 Wilkinson MG, Millar JBA. Control of the eukaryotic cell cycle by MAP kinase signalling pathways. FASEB J 2000; 14: 2147-57.
• 4 Marshall CJ. Specificity of receptor tyrosine kinase signalling: transient versus sustained extracellular signal-regulated kinase activation. Cell 1995; 80: 179-85.
• 5 Marshall CJ. MAP kinase kinase kinase, MAP kinase kinase and MAP kinase. Curr Opin Genet Dev 1994; 04: 82-89.
• 6 Ip YT, Davis RJ. Signal transduction by the cjun N-terminal kinase (JNK)—from inflammation to development. Curr Opin Cell Biol 1998; 10: 205-19.
• 7 Tournier C, Hess P, Yang DD. et al. Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. Science 2000; 288: 870-4.
• 8 New L, Han J. The p38 MAP kinase pathway and its biological function. Trend Card Med 1998; 08: 220-28.
• 9 Ono K, Han J. The p38 signal transduction pathway. Activation and function. Cell Signal 2000; 12: 1-13.
• 10 Papkoff J, Chen RH, Blenis J. et al. p42 mitogen-activated protein kinase and p90 ribosomal S6 kinase are selectively phosphorylated and activated during thrombin-induced platelet activation and aggregation. Mol Cell Biol 1994; 14: 463-72.
• 11 Kramer RM, Roberts EF, Strifler BA. et al. Thrombin induces activation of p38 MAP kinase in human platelets. J Biol Chem 1995; 270: 27395-8.
• 12 Bugaud F, Nadal-Woolbold F, Levy-Toledano S. et al. Regulation of c-jun-NH2 terminal kinase and extracellular-signal regulated kinase in human platelets. Blood 1999; 94: 3800-5.
• 13 Nakashima S, Chatani Y, Nakamura M. et al. Tyrosine phosphorylation and activation of mitogen-activated protein kinases by thrombin in human platelets: possible involvement in late arachidonic acid release. Biochem Biophys Res Commun 1994; 198: 497-503.
• 14 Aharonovitz O, Granot Y. Stimulation of mitogen-activated protein kinase and Na+/H+ exchanger in human platelets. Differential effect of phorbol ester and vasopressin. J Biol Chem 1996; 271: 16494-9.
• 15 Borsch-Haubold AG, Kramer RM, Watson SP. Cytosolic phospholipase A2 is phosphorylated in collagen- and thrombin-stimulated human platelets independent of protein kinase C and mitogen-activated protein kinase. J Biol Chem 1995; 270: 25885-92.
• 16 Borsch-Haubold AG, Kramer RM, Watson SP. Phosphorylation and activation of cytosolic phospholipase A₂ by 38-kDa mitogen-activated protein kinase in collagen-stimulated human platelets. Eur J Biochem 1997; 245: 751-9.
• 17 Li Z, Xi X, Du X. A mitogen-activated protein kinase-dependent signalling pathway in the activation of platelet integrin α_IIbβ₃ . J Biol Chem 2001; 276: 42226-32.
• 18 Nadal F, Levy-Toledano S, Grelac F. et al. Negative regulation of mitogen-activated protein kinase activation by integrin α_IIbβ₃ in platelets. J Biol Chem 1997; 272: 22381-4.
• 19 Saklatvala J, Rawlinson L, Waller RJ. et al. Role for p38 mitogen-activated protein kinase in platelet aggregation caused by collagen or a thromboxane analogue. J Biol Chem 1996; 271: 6586-9.
• 20 Hackeng CM, Relou IA, Pladet MW. et al. Early platelet activation by low density lipoprotein via p38MAP kinase. Thromb Haemost 1999; 82: 1749-56.
• 21 Borsch-Haubold AG, Kramer RM, Watson SP. Inhibition of mitogen-activated protein kinase kinase does not impair primary activation of human platelets. Biochem J 1996; 318: 207-12.
• 22 Kramer RM, Roberts EF, Um SL. et al. p38 mitogen-activated protein kinase phosphorylates cytosolic phospholipase A₂ (cPLA₂) in thrombin-stimulated platelets. Evidence that proline-directed phosphorylation is not required for mobilization of arachidonic acid by cPLA₂ . J Biol Chem 1996; 271: 27723-9.
• 23 Borsch-Haubold AG, Bartoli F, Asselin J. et al. Identification of the phosphorylation sites of cytosolic phospholipase A₂ in agonist-stimulated human platelets and HeLa cells. J Biol Chem 1998; 273: 4449-58.
• 24 Clark JD, Lin LL, Kriz RW. et al. A novel arachidonic acid-selective cytosolic PLA₂ contains a Ca(2+)-dependent translocation domain with homology to PKC and GAP. Cell 1991; 65: 1043-51.
• 25 Channon JY, Leslie CC. A calcium-dependent mechanism for associating a soluble arachidonoyl-hydrolyzing phospholipase A₂ with membrane in the macrophage cell line RAW 264.7. J Biol Chem 1990; 265: 5409-13.
• 26 Kroll MH, Harris TS, Moake JL. et al. von Willebrand factor binding to platelet GPIb initiates signals for platelet activation. J Clin Invest 1991; 88: 1568-73.
• 27 Torti M, Bertoni A, Canobbio I. et al. Rap1B and Rap2B translocation to the cytoskeleton by von Willebrand factor involves FcγIIA receptor-mediated protein tyrosine phosphorylation. J Biol Chem 1999; 274: 13690-7.
• 28 Ozaki Y, Satoh K, Yatomi Y. et al. Protein tyrosine phosphorylation in human platelets induced by interaction between glycoprotein Ib and von Willebrand factor. Biochim Biophys Acta 1995; 1243: 482-8.
• 29 Asazuma N, Ozaki Y, Satoh K. et al. Glycoprotein Ib-von Willebrand factor interactions activate tyrosine kinases in human platelets. Blood 1997; 90: 4789-98.
• 30 Sullam PM, Hyun WC, Szollosi J. et al. Physical proximity and functional interplay of the glycoprotein Ib-IX-V complex and the Fc receptor FcγRIIA on the platelet plasma membrane. J Biol Chem 1998; 273: 5331-6.
• 31 Sun B, Li J, Kambayashi J. Interaction between GPIbα and FcγIIA receptor in human platelets. Biochem Biophys Res Commun 1999; 266: 24-7.
• 32 Shrimpton CN, Borthakur G, Larrucea S. et al. Localization of the adhesion receptor glycoprotein Ib-IX-V complex to lipid rafts is required for platelet adhesion and activation. J Exp Med 2002; 196: 1057-66.
• 33 Canobbio I, Bertoni A, Lova P. et al. Platelet activation by von Willebrand factor requires coordinated signalling through thromboxane A₂ and FcγIIA receptor. J Biol Chem 2001; 276: 26022-9.
• 34 Torti M, Ramaschi G, Sinigaglia F. et al. Association of the low molecular weight GTPbinding protein rap2B with the cytoskeleton during platelet aggregation. Proc Natl Acad Sci U S A 1993; 90: 7553-7.
• 35 Canobbio I, Lova P, Sinigaglia F. et al. Proline-rich tyrosine kinase 2 and focal adhesion kinase are involved in different phases of platelet activation by VWF. Thromb Haemost 2002; 87: 509-17.
• 36 Law DA, Nannizzi-Alaimo L, Ministri K. et al. Genetic and pharmacological analysis of Syk function in α_IIbβ₃ signaling in platelets. Blood 1999; 93: 2645-52.
• 37 Wu Y, Suzuki-Inoue K, Satoh K. et al. Role of Fc receptor γ-chain in platelet glycoprotein Ibmediated signaling. Blood 2000; 97: 3836-45.
• 38 Hazan-Halevy I, Seger R, Levy R. The requirement of both extracellular regulated kinase and p38 mitogen-activated protein kinase for stimulation of cytosolic phospholipase A₂ activity by either FcγRIIA or FcγRIIIB in human neutrophils. A possible role for Pyk2 but not for the Grb2-Sos-Shc complex. J Biol Chem 2000; 275: 12416-23.
• 39 Pandey P, Avraham S, Kumar S. et al. Activation of p38 mitogen-activated protein kinase by PYK2/related adhesion focal tyrosine kinase-dependent mechanism. J Biol Chem 1999; 274: 10140-4.
• 40 Borsch-Haubold AG, Pasquet S, Watson SP. Direct inhibition of cyclooxygenase-1 and -2 by the kinase inhibitors SB 203580 and PD 98059. SB 203580 also inhibits thromboxane synthase. J Biol Chem 1998; 273: 28766-72.