Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2002 Jan 15;361(Pt 2):297–305. doi: 10.1042/0264-6021:3610297

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

Stuart J Marshall 1, Naoki Asazuma 1, Denise Best 1, Peter Wonerow 1, Gary Salmon 1, Robert K Andrews 1, Steve P Watson 1
PMCID: PMC1222309  PMID: 11772401

Abstract

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) gamma-chain and Syk, leading to tyrosine phosphorylation of phospholipase Cgamma2 (PLCgamma2). 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 gamma-chain, Syk and PLCgamma2, 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 (PP1). Under the same conditions, neither stimulus induced activation of PLCgamma2 nor functional responses, such as Ca(2+) 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 GPIbalpha in washed platelets does not promote functional responses.

Full Text

The Full Text of this article is available as a PDF (235.8 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrews R. K., Berndt M. C. Adhesion-dependent signalling and the initiation of haemostasis and thrombosis. Histol Histopathol. 1998 Jul;13(3):837–844. doi: 10.14670/HH-13.837. [DOI] [PubMed] [Google Scholar]
  2. Andrews R. K., Booth W. J., Gorman J. J., Castaldi P. A., Berndt M. C. Purification of botrocetin from Bothrops jararaca venom. Analysis of the botrocetin-mediated interaction between von Willebrand factor and the human platelet membrane glycoprotein Ib-IX complex. Biochemistry. 1989 Oct 17;28(21):8317–8326. doi: 10.1021/bi00447a009. [DOI] [PubMed] [Google Scholar]
  3. Andrews R. K., Kroll M. H., Ward C. M., Rose J. W., Scarborough R. M., Smith A. I., López J. A., Berndt M. C. Binding of a novel 50-kilodalton alboaggregin from Trimeresurus albolabris and related viper venom proteins to the platelet membrane glycoprotein Ib-IX-V complex. Effect on platelet aggregation and glycoprotein Ib-mediated platelet activation. Biochemistry. 1996 Sep 24;35(38):12629–12639. doi: 10.1021/bi960704e. [DOI] [PubMed] [Google Scholar]
  4. Andrews R. K., Shen Y., Gardiner E. E., Dong J. F., López J. A., Berndt M. C. The glycoprotein Ib-IX-V complex in platelet adhesion and signaling. Thromb Haemost. 1999 Aug;82(2):357–364. [PubMed] [Google Scholar]
  5. Asazuma N., Marshall S. J., Berlanga O., Snell D., Poole A. W., Berndt M. C., Andrews R. K., Watson S. P. The snake venom toxin alboaggregin-A activates glycoprotein VI. Blood. 2001 Jun 15;97(12):3989–3991. doi: 10.1182/blood.v97.12.3989. [DOI] [PubMed] [Google Scholar]
  6. Asazuma N., Ozaki Y., Satoh K., Yatomi Y., Handa M., Fujimura Y., Miura S., Kume S. Glycoprotein Ib-von Willebrand factor interactions activate tyrosine kinases in human platelets. Blood. 1997 Dec 15;90(12):4789–4798. [PubMed] [Google Scholar]
  7. Berndt M. C., Du X. P., Booth W. J. Ristocetin-dependent reconstitution of binding of von Willebrand factor to purified human platelet membrane glycoprotein Ib-IX complex. Biochemistry. 1988 Jan 26;27(2):633–640. doi: 10.1021/bi00402a021. [DOI] [PubMed] [Google Scholar]
  8. Canobbio I., Bertoni A., Lova P., Paganini S., Hirsch E., Sinigaglia F., Balduini C., Torti M. Platelet activation by von Willebrand factor requires coordinated signaling through thromboxane A2 and Fc gamma IIA receptor. J Biol Chem. 2001 May 8;276(28):26022–26029. doi: 10.1074/jbc.M102639200. [DOI] [PubMed] [Google Scholar]
  9. De Marco L., Girolami A., Russell S., Ruggeri Z. M. Interaction of asialo von Willebrand factor with glycoprotein Ib induces fibrinogen binding to the glycoprotein IIb/IIIa complex and mediates platelet aggregation. J Clin Invest. 1985 Apr;75(4):1198–1203. doi: 10.1172/JCI111816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dong J. F., Berndt M. C., Schade A., McIntire L. V., Andrews R. K., López J. A. Ristocetin-dependent, but not botrocetin-dependent, binding of von Willebrand factor to the platelet glycoprotein Ib-IX-V complex correlates with shear-dependent interactions. Blood. 2001 Jan 1;97(1):162–168. doi: 10.1182/blood.v97.1.162. [DOI] [PubMed] [Google Scholar]
  11. Dörmann D., Clemetson J. M., Navdaev A., Kehrel B. E., Clemetson K. J. Alboaggregin A activates platelets by a mechanism involving glycoprotein VI as well as glycoprotein Ib. Blood. 2001 Feb 15;97(4):929–936. doi: 10.1182/blood.v97.4.929. [DOI] [PubMed] [Google Scholar]
  12. Emsley J., Cruz M., Handin R., Liddington R. Crystal structure of the von Willebrand Factor A1 domain and implications for the binding of platelet glycoprotein Ib. J Biol Chem. 1998 Apr 24;273(17):10396–10401. doi: 10.1074/jbc.273.17.10396. [DOI] [PubMed] [Google Scholar]
  13. Englund G. D., Bodnar R. J., Li Z., Ruggeri Z. M., Du X. Regulation of von Willebrand factor binding to the platelet glycoprotein Ib-IX by a membrane skeleton-dependent inside-out signal. J Biol Chem. 2001 Feb 26;276(20):16952–16959. doi: 10.1074/jbc.M008048200. [DOI] [PubMed] [Google Scholar]
  14. Escolar G., Monteagudo J., Villamor N., Garrido M., Castillo R., Ordinas A. Ristocetin induces platelet aggregation: a morphological demonstration. Br J Haematol. 1988 Jul;69(3):379–386. doi: 10.1111/j.1365-2141.1988.tb02377.x. [DOI] [PubMed] [Google Scholar]
  15. Francesconi M., Casonato A., Pontara E., Dalla Via L., Girolami A., Deana R. Type IIB von Willebrand factor induces phospholipase A2 activation and cytosolic Ca2+ increase in platelets. Biochem Biophys Res Commun. 1995 Sep 5;214(1):102–109. doi: 10.1006/bbrc.1995.2262. [DOI] [PubMed] [Google Scholar]
  16. Francischetti I. M., Saliou B., Leduc M., Carlini C. R., Hatmi M., Randon J., Faili A., Bon C. Convulxin, a potent platelet-aggregating protein from Crotalus durissus terrificus venom, specifically binds to platelets. Toxicon. 1997 Aug;35(8):1217–1228. doi: 10.1016/s0041-0101(97)00021-4. [DOI] [PubMed] [Google Scholar]
  17. Fujimura Y., Kawasaki T., Titani K. Snake venom proteins modulating the interaction between von Willebrand factor and platelet glycoprotein Ib. Thromb Haemost. 1996 Nov;76(5):633–639. [PubMed] [Google Scholar]
  18. Gibbins J., Asselin J., Farndale R., Barnes M., Law C. L., Watson S. P. Tyrosine phosphorylation of the Fc receptor gamma-chain in collagen-stimulated platelets. J Biol Chem. 1996 Jul 26;271(30):18095–18099. doi: 10.1074/jbc.271.30.18095. [DOI] [PubMed] [Google Scholar]
  19. Giles H., Leff P., Bolofo M. L., Kelly M. G., Robertson A. D. The classification of prostaglandin DP-receptors in platelets and vasculature using BW A868C, a novel, selective and potent competitive antagonist. Br J Pharmacol. 1989 Feb;96(2):291–300. doi: 10.1111/j.1476-5381.1989.tb11816.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Grainick H. R., Williams S. B., Coller B. S. Asialo von Willebrand factor interactions with platelets. Interdependence of glycoproteins Ib and IIb/IIIa for binding and aggregation. J Clin Invest. 1985 Jan;75(1):19–25. doi: 10.1172/JCI111673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gross B. S., Lee J. R., Clements J. L., Turner M., Tybulewicz V. L., Findell P. R., Koretzky G. A., Watson S. P. Tyrosine phosphorylation of SLP-76 is downstream of Syk following stimulation of the collagen receptor in platelets. J Biol Chem. 1999 Feb 26;274(9):5963–5971. doi: 10.1074/jbc.274.9.5963. [DOI] [PubMed] [Google Scholar]
  22. Haneline L. S., Gobbett T. A., Ramani R., Carreau M., Buchwald M., Yoder M. C., Clapp D. W. Loss of FancC function results in decreased hematopoietic stem cell repopulating ability. Blood. 1999 Jul 1;94(1):1–8. [PubMed] [Google Scholar]
  23. Ikeda Y., Handa M., Kawano K., Kamata T., Murata M., Araki Y., Anbo H., Kawai Y., Watanabe K., Itagaki I. The role of von Willebrand factor and fibrinogen in platelet aggregation under varying shear stress. J Clin Invest. 1991 Apr;87(4):1234–1240. doi: 10.1172/JCI115124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jackson S. P., Schoenwaelder S. M., Yuan Y., Rabinowitz I., Salem H. H., Mitchell C. A. Adhesion receptor activation of phosphatidylinositol 3-kinase. von Willebrand factor stimulates the cytoskeletal association and activation of phosphatidylinositol 3-kinase and pp60c-src in human platelets. J Biol Chem. 1994 Oct 28;269(43):27093–27099. [PubMed] [Google Scholar]
  25. Janes P. W., Ley S. C., Magee A. I., Kabouridis P. S. The role of lipid rafts in T cell antigen receptor (TCR) signalling. Semin Immunol. 2000 Feb;12(1):23–34. doi: 10.1006/smim.2000.0204. [DOI] [PubMed] [Google Scholar]
  26. Kowalska M. A., Tan L., Holt J. C., Peng M., Karczewski J., Calvete J. J., Niewiarowski S. Alboaggregins A and B. Structure and interaction with human platelets. Thromb Haemost. 1998 Mar;79(3):609–613. [PubMed] [Google Scholar]
  27. Kroll M. H., Harris T. S., Moake J. L., Handin R. I., Schafer A. I. von Willebrand factor binding to platelet GpIb initiates signals for platelet activation. J Clin Invest. 1991 Nov;88(5):1568–1573. doi: 10.1172/JCI115468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kroll M. H., Hellums J. D., Guo Z., Durante W., Razdan K., Hrbolich J. K., Schafer A. I. Protein kinase C is activated in platelets subjected to pathological shear stress. J Biol Chem. 1993 Feb 15;268(5):3520–3524. [PubMed] [Google Scholar]
  29. Kroll M. H., Hellums J. D., McIntire L. V., Schafer A. I., Moake J. L. Platelets and shear stress. Blood. 1996 Sep 1;88(5):1525–1541. [PubMed] [Google Scholar]
  30. López J. A. The platelet glycoprotein Ib-IX complex. Blood Coagul Fibrinolysis. 1994 Feb;5(1):97–119. [PubMed] [Google Scholar]
  31. Milner E. P., Zheng Q., Kermode J. C. Ristocetin-mediated interaction of human von Willebrand factor with platelet glycoprotein lb evokes a transient calcium signal: observations with Fura-PE3. J Lab Clin Med. 1998 Jan;131(1):49–62. doi: 10.1016/s0022-2143(98)90077-4. [DOI] [PubMed] [Google Scholar]
  32. Navdaev A., Dörmann D., Clemetson J. M., Clemetson K. J. Echicetin, a GPIb-binding snake C-type lectin from Echis carinatus, also contains a binding site for IgMkappa responsible for platelet agglutination in plasma and inducing signal transduction. Blood. 2001 Apr 15;97(8):2333–2341. doi: 10.1182/blood.v97.8.2333. [DOI] [PubMed] [Google Scholar]
  33. Pasquet J. M., Gross B., Quek L., Asazuma N., Zhang W., Sommers C. L., Schweighoffer E., Tybulewicz V., Judd B., Lee J. R. LAT is required for tyrosine phosphorylation of phospholipase cgamma2 and platelet activation by the collagen receptor GPVI. Mol Cell Biol. 1999 Dec;19(12):8326–8334. doi: 10.1128/mcb.19.12.8326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Peng M., Lu W., Beviglia L., Niewiarowski S., Kirby E. P. Echicetin: a snake venom protein that inhibits binding of von Willebrand factor and alboaggregins to platelet glycoprotein Ib. Blood. 1993 May 1;81(9):2321–2328. [PubMed] [Google Scholar]
  35. Read M. S., Smith S. V., Lamb M. A., Brinkhous K. M. Role of botrocetin in platelet agglutination: formation of an activated complex of botrocetin and von Willebrand factor. Blood. 1989 Aug 15;74(3):1031–1035. [PubMed] [Google Scholar]
  36. Ruggeri Z. M. Structure and function of von Willebrand factor. Thromb Haemost. 1999 Aug;82(2):576–584. [PubMed] [Google Scholar]
  37. Ruggeri Z. M., Zimmerman T. S. The complex multimeric composition of factor VIII/von Willebrand factor. Blood. 1981 Jun;57(6):1140–1143. [PubMed] [Google Scholar]
  38. Ruggeri Z. M., Zimmerman T. S. von Willebrand factor and von Willebrand disease. Blood. 1987 Oct;70(4):895–904. [PubMed] [Google Scholar]
  39. Sadler J. E. Biochemistry and genetics of von Willebrand factor. Annu Rev Biochem. 1998;67:395–424. doi: 10.1146/annurev.biochem.67.1.395. [DOI] [PubMed] [Google Scholar]
  40. Satoh K., Asazuma N., Yatomi Y., Fujimura Y., Miura S., Titani K., Ozaki Y. Activation of protein-tyrosine kinase pathways in human platelets stimulated with the A1 domain of von Willebrand factor. Platelets. 2000 May;11(3):171–176. doi: 10.1080/095371000403116. [DOI] [PubMed] [Google Scholar]
  41. Shen Y., Romo G. M., Dong J. F., Schade A., McIntire L. V., Kenny D., Whisstock J. C., Berndt M. C., López J. A., Andrews R. K. Requirement of leucine-rich repeats of glycoprotein (GP) Ibalpha for shear-dependent and static binding of von Willebrand factor to the platelet membrane GP Ib-IX-V complex. Blood. 2000 Feb 1;95(3):903–910. [PubMed] [Google Scholar]
  42. Simons K., Toomre D. Lipid rafts and signal transduction. Nat Rev Mol Cell Biol. 2000 Oct;1(1):31–39. doi: 10.1038/35036052. [DOI] [PubMed] [Google Scholar]
  43. Weiss H. J. Abnormalities of factor VIII and platelet aggregation--use of ristocetin in diagnosing the von Willebrand syndrome. Blood. 1975 Mar;45(3):403–412. [PubMed] [Google Scholar]
  44. Weiss H. J., Rogers J., Brand H. Defective ristocetin-induced platelet aggregation in von Willebrand's disease and its correction by factor VIII. J Clin Invest. 1973 Nov;52(11):2697–2707. doi: 10.1172/JCI107464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wu Y., Suzuki-Inoue K., Satoh K., Asazuma N., Yatomi Y., Berndt M. C., Ozaki Y. Role of Fc receptor gamma-chain in platelet glycoprotein Ib-mediated signaling. Blood. 2001 Jun 15;97(12):3836–3845. doi: 10.1182/blood.v97.12.3836. [DOI] [PubMed] [Google Scholar]
  46. Yanabu M., Ozaki Y., Nomura S., Miyake T., Miyazaki Y., Kagawa H., Yamanaka Y., Asazuma N., Satoh K., Kume S. Tyrosine phosphorylation and p72syk activation by an anti-glycoprotein Ib monoclonal antibody. Blood. 1997 Mar 1;89(5):1590–1598. [PubMed] [Google Scholar]
  47. Yanaga F., Poole A., Asselin J., Blake R., Schieven G. L., Clark E. A., Law C. L., Watson S. P. Syk interacts with tyrosine-phosphorylated proteins in human platelets activated by collagen and cross-linking of the Fc gamma-IIA receptor. Biochem J. 1995 Oct 15;311(Pt 2):471–478. doi: 10.1042/bj3110471. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES