Thromb Haemost 2003; 89(06): 1016-1023
DOI: 10.1055/s-0037-1613403
Platelets and Blood Cells
Schattauer GmbH

Platelet P-selectin expression: requirement for protein kinase C, but not protein tyrosine kinase or phosphoinositide 3-kinase

Danielle Libersan
1   Montreal Heart Institute and the University of Montreal, Montreal, Quebec, Canada
,
Yahye Merhi
1   Montreal Heart Institute and the University of Montreal, Montreal, Quebec, Canada
› Author Affiliations
Financial support: This study was supported by grants from the Canadian Institutes of Health Research and the Heart and Stroke Foundation of Quebec.
Further Information

Publication History

Received 22 October 2002

Accepted after revision 30 January 2003

Publication Date:
08 December 2017 (online)

Summary

P-selectin is translocated from the α-granules to the surface of activated platelets where it participates in thrombosis and inflammation. We investigated the signaling pathways involved in thrombin-induced human platelet P-selectin expression. Assessed by flow cytometry, inhibition of protein kinase C (PKC) with chelerythrine reduced P-selectin expression by 66%, platelet/neutrophil binding, GPIIb/IIIa activation and aggregation (p<0.05). Gö 6976, an inhibitor of the conventional PKCs (α and β), did not alter P-selectin expression. However, rottlerin inhibited by 50% its expression (p<0.05), but only at doses that interfere with the novel (є, η) and atypical (ζ) PKCs. Inhibition of protein tyrosine kinase (PTK) and phosphoinositide 3-kinase (PI3-K) did not significantly affect P-selectin expression. In conclusion, thrombin-induced P-selectin expression is PKC-sensitive, but PTK and PI3-K-insensitive. The novel є and η and atypical ζ, but not the conventional α and β and the novel θ PKCs, may be involved in this process.

 
  • References

  • 1 Berman CL, Yeo EL, Wencel-Drake JD, Furie BC, Ginsberg MH, Furie B. A platelet alpha granule membrane protein that is associated with the plasma membrane after activation. Characterization and subcellular localization of platelet activation-dependent granule-external membrane protein. J Clin Invest 1986; 78: 130-7.
  • 2 McEver RP, Beckstead JH, Moore KL, Marshall-Carlson L, Bainton DF. GMP-140, a platelet α-granule membrane protein, is also synthetized by vascular endothelial cells and is localized in Weibel-Palade bodies. J Clin Invest 1989; 84: 92-9.
  • 3 Yang J, Furie BC, Furie B. The biology of P-selectin glycoprotein ligand-1: its role as a selectin counterreceptor in leukocyte-endothelial and leukocyte-platelet interaction. Thromb Haemost 1999; 81: 1-7.
  • 4 Frenette PS, Johnson RC, Hynes RO, Wagner DD. Platelets roll on stimulated endothelium in vivo: an interaction mediated by endothelial P-selectin. Proc Natl Acad Sci USA 1995; 92: 7450-4.
  • 5 Frenette PS, Moyna C, Hartwell DW, Lowe JB, Hynes RO, Wagner DD. Platelet-endothelial interactions in inflammed mesenteric venules. Blood 1998; 91: 1318-24.
  • 6 Merten M, Thiagarajan P. P-selectin expression on platelets determines size and stability of platelet aggregates. Circulation 2000; 102: 1931-6.
  • 7 Brass LF, Manning DR, Cichowski K, Abrams CS. Signaling through G proteins in platelets: to the integrins and beyond. Thromb Haemost 1997; 78: 581-9.
  • 8 Wang F, Naik UP, Ehrlich YH, Freyberg Z, Osada S, Ohno S, Kuroki T, Suzuki K, Kornecki E. A new protein kinase C, nPKC eta’, and nPKC theta are expressed in human platelets: involvement of nPKC eta’ and nPKC theta in signal transduction stimulated by PAF. Biochem Biophys Res Commun 1993; 191: 240-6.
  • 9 Gerrard JM, Beattie LL, Park J, Israels SJ, McNicol A, Lint D, Cragoe Jr EJ. A role for protein kinase C in the membrane fusion necessary for platelet granule secretion. Blood 1989; 74: 2405-13.
  • 10 Geanacopoulos M, Turner J, Bowling KE, Vandenberg SR, Gear AR. The role of protein kinase C in the initial events of platelet activation by thrombin assessed with a selective inhibitor. Thromb Res 1993; 69: 113-24.
  • 11 Walker TR, Watson SP. Synergy between Ca2+ and protein kinase C is the major factor in determining the level of secretion from human platelets. Biochem J 1993; 289: 277-82.
  • 12 Chung S-H, Polgár J, Reed GL. Protein kinase C phosphorylation of syntaxin 4 in thrombin-activated human platelets. J Biol Chem 2000; 275: 25286-91.
  • 13 Chen M, Geng J-G. Inhibition of protein tyrosine phosphatases suppresses P-selectin exocytosis in activated human platelets. Biochem Biophys Res Commun 2001; 286: 831-8.
  • 14 Kovacsovics TJ, Bachelot C, Toker A, Vlahos CJ, Duckworth B, Cantley LC, Hartwig JH. Phosphoinositide 3-kinase inhibition spares actin assembly in activating platelets but reverses platelet aggregation. J Biol Chem 1995; 270: 11358-66.
  • 15 Rozenvayn N, Flaumenhaft R. Phosphatidylinositol 4,5-biphosphate mediates Ca2+-induced platelet α-granule secretion. Evidence for type II phosphatidylinositol 5-phosphate 4-kinase function. J Biol Chem 2001; 276: 22410-9.
  • 16 Théorêt J-F, Bienvenu J-G, Kumar A, Merhi Y. P-selectin antagonism with recombinant P-selectin glycoprotein ligand-1 (rPSGL-1) inhibits circulating activated platelet binding to neutrophils induced by damaged arterial surfaces. J Pharmacol Exp Ther 2001; 298: 658-64.
  • 17 Libersan D, Rousseau G, Merhi Y. Differential regulation of P-selectin expression by protein kinase A and protein kinase G in thrombin-stimulated human platelets. Thromb Haemost 2003; 89: 10-7.
  • 18 Rittenhouse SE. Phosphoinositide 3-kinase activation and platelet function. Blood 1996; 88: 4401-14.
  • 19 Lévy-Toledano S, Gallet C, Nadal F, Bryckaert M, Maclouf J, Rosa JP. Phosphorylation and dephosphorylation mechanisms in platelet function: a tightly regulated balance. Thromb Haemost 1997; 78: 226-33.
  • 20 Santos MT, Moscardó A, Vallés J, Martínez M, Piñón M, Aznar J, Broekman MJ, Marcus AJ. Participation of tyrosine phosphorylation in cytoskeletal reorganization, αIIbβ3 integrin receptor activation, and aspirin-insensitive mechanisms of thrombin-stimulated human platelets. Circulation 2000; 102: 1924-30.
  • 21 Shebuski RJ, Kilgore KS. Role of inflammatory mediators in thrombogenesis. J Pharmacol Exp Ther 2002; 300: 729-35.
  • 22 Handa K, Igarashi Y, Nisar M, Hakomori S-I. Downregulation of GMP-140 (CD62 or PADGEM) expression on platelets by N, N-dimethyl and N, N, N-trimethyl derivatives of sphingosine. Biochemistry 1991; 30: 11682-6.
  • 23 Murohara T, Parkinson SJ, Waldman SA, Lefer AM. Inhibition of nitric oxide biosyn-thesis promotes P-selectin expression in platelets. Role of protein kinase C. Arterioscler Thromb Vasc Biol 1995; 15: 2068-75.
  • 24 Murohara T, Scalia R, Lefer AM. Lysophosphatidylcholine promotes P-selectin expression in platelets and endothelial cells. Possible involvement of protein kinase C activation and its inhibition by nitric oxide donors. Circ Res 1996; 78: 780-9.
  • 25 Lévy-Toledano S. Platelet signal transduction pathways: could we organize them into a ‘hierarchy’?. Haemostasis 1999; 29: 4-15.
  • 26 Jackson SP, Schoenwaelder SM, Yuan Y, Salem HH, Cooray P. Non-receptor protein tyrosine kinases and phosphatases in human platelets. Thromb Res 1996; 76: 640-50.
  • 27 Caron A, Théoret J-F, Mousa SA, Merhi Y. Anti-platelet effects of GPIIb/IIIa and P-selectin antagonism, platelet activation, and binding to neutrophils. J Cardiovasc Pharma-col 2002; 40: 296-306.
  • 28 Selheim F, Holmsen H, Vassbotn FS. PI 3-kinase signalling in platelets: the significance of synergistic, autocrine stimulation. Platelets 2000; 11: 69-82.
  • 29 Pasquet J-M, Gross BS, Gratacap M-P, Quek L, Pasquet S, Payrastre B, van Willigen G, Mountford JC, Watson SP. Thrombopoietin potentiates collagen receptor signaling in platelets through a phosphatidylinositol 3-kinase-dependent pathway. Blood 2000; 95: 3429-34.