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. 1993 Jul;92(1):288–296. doi: 10.1172/JCI116563

Thrombospondin-platelet interactions. Role of divalent cations, wall shear rate, and platelet membrane glycoproteins.

F R Agbanyo 1, J J Sixma 1, P G de Groot 1, L R Languino 1, E F Plow 1
PMCID: PMC293590  PMID: 7686921

Abstract

The role of thrombospondin, a multifunctional matrix glycoprotein, in platelet adhesion is controversial: both adhesive and antiadhesive properties have been attributed to this molecule. Because shear flow has a significant influence on platelet adhesion, we have assessed thrombospondin-platelet interactions both under static and flow conditions. The capacity of thrombospondin to support platelet adhesion depended upon its conformation. In a Ca(2+)-depleted conformation, such as in citrated plasma, thrombospondin was nonadhesive or antiadhesive as it inhibited platelet adhesion to fibrinogen, fibronectin, laminin, and von Willebrand factor by 30-70%. In a Ca(2+)-replete conformation, however, thrombospondin effectively supported platelet adhesion. Shear rate influenced this adhesion; percent surface coverage on thrombospondin increased from 5.4 +/- 0.3 at 0 s-1 to 41.5 +/- 6.7 at 1,600 s-1. In contrast to the extensive platelet spreading observed on fibronectin at all shear rates, platelet spreading on thrombospondin occurred only sporadically and at high shear rates. GPIa-IIa, GPIIb-IIIa, GPIV, and the vitronectin receptor, which are all proposed platelet receptors for thrombospondin, were not solely responsible for platelet adhesion to thrombospondin. These results suggest that thrombospondin may play a dual role in adhesive processes in vivo: (a) it may function in conjunction with other adhesive proteins to maintain optimal platelet adhesion at various shear rates; and (b) it may serve as a modulator of cellular adhesive functions under specific microenvironmental conditions.

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Selected References

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  1. Aiken M. L., Ginsberg M. H., Byers-Ward V., Plow E. F. Effects of OKM5, a monoclonal antibody to glycoprotein IV, on platelet aggregation and thrombospondin surface expression. Blood. 1990 Dec 15;76(12):2501–2509. [PubMed] [Google Scholar]
  2. Aiken M. L., Ginsberg M. H., Plow E. F. Divalent cation-dependent and independent surface expression of thrombospondin on thrombin-stimulated human platelets. Blood. 1987 Jan;69(1):58–64. [PubMed] [Google Scholar]
  3. Asch A. S., Barnwell J., Silverstein R. L., Nachman R. L. Isolation of the thrombospondin membrane receptor. J Clin Invest. 1987 Apr;79(4):1054–1061. doi: 10.1172/JCI112918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baenziger N. L., Brodie G. N., Majerus P. W. Isolation and properties of a thrombin-sensitive protein of human platelets. J Biol Chem. 1972 May 10;247(9):2723–2731. [PubMed] [Google Scholar]
  5. Baumgartner H. R., Muggli R., Tschopp T. B., Turitto V. T. Platelet adhesion, release and aggregation in flowing blood: effects of surface properties and platelet function. Thromb Haemost. 1976 Feb 29;35(1):124–138. [PubMed] [Google Scholar]
  6. Baumgartner H. R., Turitto V., Weiss H. J. Effect of shear rate on platelet interaction with subendothelium in citrated and native blood. II. Relationships among platelet adhesion, thrombus dimensions, and fibrin formation. J Lab Clin Med. 1980 Feb;95(2):208–221. [PubMed] [Google Scholar]
  7. Chen K., Lin Y., Detwiler T. C. Protein disulfide isomerase activity is released by activated platelets. Blood. 1992 May 1;79(9):2226–2228. [PubMed] [Google Scholar]
  8. Clezardin P., Jouishomme H., Chavassieux P., Marie P. J. Thrombospondin is synthesized and secreted by human osteoblasts and osteosarcoma cells. A model to study the different effects of thrombospondin in cell adhesion. Eur J Biochem. 1989 May 15;181(3):721–726. doi: 10.1111/j.1432-1033.1989.tb14783.x. [DOI] [PubMed] [Google Scholar]
  9. Coller B. S., Beer J. H., Scudder L. E., Steinberg M. H. Collagen-platelet interactions: evidence for a direct interaction of collagen with platelet GPIa/IIa and an indirect interaction with platelet GPIIb/IIIa mediated by adhesive proteins. Blood. 1989 Jul;74(1):182–192. [PubMed] [Google Scholar]
  10. Dixit V. M., Galvin N. J., O'Rourke K. M., Frazier W. A. Monoclonal antibodies that recognize calcium-dependent structures of human thrombospondin. Characterization and mapping of their epitopes. J Biol Chem. 1986 Feb 5;261(4):1962–1968. [PubMed] [Google Scholar]
  11. Galvin N. J., Vance P. M., Dixit V. M., Fink B., Frazier W. A. Interaction of human thrombospondin with types I-V collagen: direct binding and electron microscopy. J Cell Biol. 1987 May;104(5):1413–1422. doi: 10.1083/jcb.104.5.1413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hantgan R. R., Hindriks G., Taylor R. G., Sixma J. J., de Groot P. G. Glycoprotein Ib, von Willebrand factor, and glycoprotein IIb:IIIa are all involved in platelet adhesion to fibrin in flowing whole blood. Blood. 1990 Jul 15;76(2):345–353. [PubMed] [Google Scholar]
  13. Houdijk W. P., Sakariassen K. S., Nievelstein P. F., Sixma J. J. Role of factor VIII-von Willebrand factor and fibronectin in the interaction of platelets in flowing blood with monomeric and fibrillar human collagen types I and III. J Clin Invest. 1985 Feb;75(2):531–540. doi: 10.1172/JCI111729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Houdijk W. P., de Groot P. G., Nievelstein P. F., Sakariassen K. S., Sixma J. J. Subendothelial proteins and platelet adhesion. von Willebrand factor and fibronectin, not thrombospondin, are involved in platelet adhesion to extracellular matrix of human vascular endothelial cells. Arteriosclerosis. 1986 Jan-Feb;6(1):24–33. doi: 10.1161/01.atv.6.1.24. [DOI] [PubMed] [Google Scholar]
  15. Karczewski J., Knudsen K. A., Smith L., Murphy A., Rothman V. L., Tuszynski G. P. The interaction of thrombospondin with platelet glycoprotein GPIIb-IIIa. J Biol Chem. 1989 Dec 15;264(35):21322–21326. [PubMed] [Google Scholar]
  16. Kehrel B., Balleisen L., Kokott R., Mesters R., Stenzinger W., Clemetson K. J., van de Loo J. Deficiency of intact thrombospondin and membrane glycoprotein Ia in platelets with defective collagen-induced aggregation and spontaneous loss of disorder. Blood. 1988 Apr;71(4):1074–1078. [PubMed] [Google Scholar]
  17. Kehrel B., Kronenberg A., Schwippert B., Niesing-Bresch D., Niehues U., Tschöpe D., van de Loo J., Clemetson K. J. Thrombospondin binds normally to glycoprotein IIIb deficient platelets. Biochem Biophys Res Commun. 1991 Sep 16;179(2):985–991. doi: 10.1016/0006-291x(91)91915-y. [DOI] [PubMed] [Google Scholar]
  18. Kirchhofer D., Languino L. R., Ruoslahti E., Pierschbacher M. D. Alpha 2 beta 1 integrins from different cell types show different binding specificities. J Biol Chem. 1990 Jan 15;265(2):615–618. [PubMed] [Google Scholar]
  19. Lahav J., Lawler J., Gimbrone M. A. Thrombospondin interactions with fibronectin and fibrinogen. Mutual inhibition in binding. Eur J Biochem. 1984 Nov 15;145(1):151–156. doi: 10.1111/j.1432-1033.1984.tb08534.x. [DOI] [PubMed] [Google Scholar]
  20. Lahav J. Thrombospondin inhibits adhesion of endothelial cells. Exp Cell Res. 1988 Jul;177(1):199–204. doi: 10.1016/0014-4827(88)90037-7. [DOI] [PubMed] [Google Scholar]
  21. Lahav J. Thrombospondin inhibits adhesion of platelets to glass and protein-covered substrata. Blood. 1988 Apr;71(4):1096–1099. [PubMed] [Google Scholar]
  22. Lawler J., Chao F. C., Cohen C. M. Evidence for calcium-sensitive structure in platelet thrombospondin. Isolation and partial characterization of thrombospondin in the presence of calcium. J Biol Chem. 1982 Oct 25;257(20):12257–12265. [PubMed] [Google Scholar]
  23. Lawler J., Hynes R. O. An integrin receptor on normal and thrombasthenic platelets that binds thrombospondin. Blood. 1989 Nov 1;74(6):2022–2027. [PubMed] [Google Scholar]
  24. Lawler J., Simons E. R. Cooperative binding of calcium to thrombospondin. The effect of calcium on the circular dichroism and limited tryptic digestion of thrombospondin. J Biol Chem. 1983 Oct 25;258(20):12098–12101. [PubMed] [Google Scholar]
  25. Lawler J., Weinstein R., Hynes R. O. Cell attachment to thrombospondin: the role of ARG-GLY-ASP, calcium, and integrin receptors. J Cell Biol. 1988 Dec;107(6 Pt 1):2351–2361. doi: 10.1083/jcb.107.6.2351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Leung L. L., Li W. X., McGregor J. L., Albrecht G., Howard R. J. CD36 peptides enhance or inhibit CD36-thrombospondin binding. A two-step process of ligand-receptor interaction. J Biol Chem. 1992 Sep 5;267(25):18244–18250. [PubMed] [Google Scholar]
  27. Leung L. L., Nachman R. L. Complex formation of platelet thrombospondin with fibrinogen. J Clin Invest. 1982 Sep;70(3):542–549. doi: 10.1172/JCI110646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Leung L. L. Role of thrombospondin in platelet aggregation. J Clin Invest. 1984 Nov;74(5):1764–1772. doi: 10.1172/JCI111595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. McGregor J. L., Catimel B., Parmentier S., Clezardin P., Dechavanne M., Leung L. L. Rapid purification and partial characterization of human platelet glycoprotein IIIb. Interaction with thrombospondin and its role in platelet aggregation. J Biol Chem. 1989 Jan 5;264(1):501–506. [PubMed] [Google Scholar]
  30. Mosher D. F., Doyle M. J., Jaffe E. A. Synthesis and secretion of thrombospondin by cultured human endothelial cells. J Cell Biol. 1982 May;93(2):343–348. doi: 10.1083/jcb.93.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mumby S. M., Raugi G. J., Bornstein P. Interactions of thrombospondin with extracellular matrix proteins: selective binding to type V collagen. J Cell Biol. 1984 Feb;98(2):646–652. doi: 10.1083/jcb.98.2.646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Murphy-Ullrich J. E. A different approach to determining platelet thrombospondin receptors. J Lab Clin Med. 1991 Nov;118(5):418–420. [PubMed] [Google Scholar]
  33. Murphy-Ullrich J. E., Hök M. Thrombospondin modulates focal adhesions in endothelial cells. J Cell Biol. 1989 Sep;109(3):1309–1319. doi: 10.1083/jcb.109.3.1309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pytela R., Pierschbacher M. D., Argraves S., Suzuki S., Ruoslahti E. Arginine-glycine-aspartic acid adhesion receptors. Methods Enzymol. 1987;144:475–489. doi: 10.1016/0076-6879(87)44196-7. [DOI] [PubMed] [Google Scholar]
  35. Pytela R., Pierschbacher M. D., Ginsberg M. H., Plow E. F., Ruoslahti E. Platelet membrane glycoprotein IIb/IIIa: member of a family of Arg-Gly-Asp--specific adhesion receptors. Science. 1986 Mar 28;231(4745):1559–1562. doi: 10.1126/science.2420006. [DOI] [PubMed] [Google Scholar]
  36. Raugi G. J., Mumby S. M., Abbott-Brown D., Bornstein P. Thrombospondin: synthesis and secretion by cells in culture. J Cell Biol. 1982 Oct;95(1):351–354. doi: 10.1083/jcb.95.1.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sakariassen K. S., Aarts P. A., de Groot P. G., Houdijk W. P., Sixma J. J. A perfusion chamber developed to investigate platelet interaction in flowing blood with human vessel wall cells, their extracellular matrix, and purified components. J Lab Clin Med. 1983 Oct;102(4):522–535. [PubMed] [Google Scholar]
  38. Sakariassen K. S., Fressinaud E., Girma J. P., Baumgartner H. R., Meyer D. Mediation of platelet adhesion to fibrillar collagen in flowing blood by a proteolytic fragment of human von Willebrand factor. Blood. 1986 May;67(5):1515–1518. [PubMed] [Google Scholar]
  39. Santoro S. A., Rajpara S. M., Staatz W. D., Woods V. L., Jr Isolation and characterization of a platelet surface collagen binding complex related to VLA-2. Biochem Biophys Res Commun. 1988 May 31;153(1):217–223. doi: 10.1016/s0006-291x(88)81211-7. [DOI] [PubMed] [Google Scholar]
  40. Slane J. M., Mosher D. F., Lai C. S. Conformational change in thrombospondin induced by removal of bound Ca2+. A spin label approach. FEBS Lett. 1988 Mar 14;229(2):363–366. doi: 10.1016/0014-5793(88)81157-8. [DOI] [PubMed] [Google Scholar]
  41. Speziale M. V., Detwiler T. C. Free thiols of platelet thrombospondin. Evidence for disulfide isomerization. J Biol Chem. 1990 Oct 15;265(29):17859–17867. [PubMed] [Google Scholar]
  42. Stel H. V., Sakariassen K. S., de Groot P. G., van Mourik J. A., Sixma J. J. Von Willebrand factor in the vessel wall mediates platelet adherence. Blood. 1985 Jan;65(1):85–90. [PubMed] [Google Scholar]
  43. Sun X., Skorstengaard K., Mosher D. F. Disulfides modulate RGD-inhibitable cell adhesive activity of thrombospondin. J Cell Biol. 1992 Aug;118(3):693–701. doi: 10.1083/jcb.118.3.693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tandon N. N., Ockenhouse C. F., Greco N. J., Jamieson G. A. Adhesive functions of platelets lacking glycoprotein IV (CD36). Blood. 1991 Dec 1;78(11):2809–2813. [PubMed] [Google Scholar]
  45. Tuszynski G. P., Kowalska M. A. Thrombospondin-induced adhesion of human platelets. J Clin Invest. 1991 Apr;87(4):1387–1394. doi: 10.1172/JCI115144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tuszynski G. P., Rothman V. L., Murphy A., Siegler K., Knudsen K. A. Thrombospondin promotes platelet aggregation. Blood. 1988 Jul;72(1):109–115. [PubMed] [Google Scholar]
  47. Tuszynski G. P., Rothman V., Murphy A., Siegler K., Smith L., Smith S., Karczewski J., Knudsen K. A. Thrombospondin promotes cell-substratum adhesion. Science. 1987 Jun 19;236(4808):1570–1573. doi: 10.1126/science.2438772. [DOI] [PubMed] [Google Scholar]
  48. Vuillard L., Clezardin P., Miller A. Models of human platelet thrombospondin in solution. A dynamic light-scattering study. Biochem J. 1991 Apr 1;275(Pt 1):263–266. doi: 10.1042/bj2750263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Weiss H. J., Turitto V. T., Baumgartner H. R. Further evidence that glycoprotein IIb-IIIa mediates platelet spreading on subendothelium. Thromb Haemost. 1991 Feb 12;65(2):202–205. [PubMed] [Google Scholar]
  50. Wencel-Drake J. D., Plow E. F., Zimmerman T. S., Painter R. G., Ginsberg M. H. Immunofluorescent localization of adhesive glycoproteins in resting and thrombin-stimulated platelets. Am J Pathol. 1984 May;115(2):156–164. [PMC free article] [PubMed] [Google Scholar]
  51. Willems C., Astaldi G. C., De Groot P. G., Janssen M. C., Gonsalvez M. D., Zeijlemaker W. P., Van Mourik J. A., Van Aken W. G. Media conditioned by cultured human vascular endothelial cells inhibit the growth of vascular smooth muscle cells. Exp Cell Res. 1982 May;139(1):191–197. doi: 10.1016/0014-4827(82)90332-9. [DOI] [PubMed] [Google Scholar]
  52. Wolff R., Plow E. F., Ginsberg M. H. Interaction of thrombospondin with resting and stimulated human platelets. J Biol Chem. 1986 May 25;261(15):6840–6846. [PubMed] [Google Scholar]

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