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. 1988 Jul;82(1):65–73. doi: 10.1172/JCI113602

Evidence that the primary binding site of von Willebrand factor that mediates platelet adhesion on subendothelium is not collagen.

P G de Groot 1, M Ottenhof-Rovers 1, J A van Mourik 1, J J Sixma 1
PMCID: PMC303477  PMID: 2839553

Abstract

We have studied the binding of von Willebrand factor to extracellular matrices of endothelial cells and to the vessel wall of human umbilical arteries in relation to its function in supporting platelet adhesion. CLB-RAg 201, an MAb against von Willebrand factor, completely inhibits the binding of von Willebrand factor to collagen type I and type III. CLB-RAg 201 does not inhibit the binding of 125I-von Willebrand factor to extracellular matrices of endothelial cells, to smooth muscle cells, or to the subendothelium. CLB-RAg 201 partly inhibits platelet adhesion to these surfaces, but this directly affects the interaction between von Willebrand factor and platelets and is not due to inhibition of binding of von Willebrand factor to these surfaces. Another MAb, CLB-RAg 38, does not inhibit the binding of von Willebrand factor to collagen. CLB-RAg 38 completely inhibits the binding of von Willebrand factor to extracellular matrices. CLB-RAg 38 inhibits platelet adhesion to cellular matrices completely insofar as it is dependent on plasma von Willebrand factor. CLB-RAg 38 does not inhibit the total binding of von Willebrand factor to subendothelium, as there are too many different binding sites, but it completely inhibits the functional binding sites for von Willebrand factor that support platelet adhesion. The epitopes for CLB-RAg 38 and 201 on the von Willebrand factor molecule are located on different fragments of the molecule. These results indicate that von Willebrand factor binds to subendothelium and matrices of cultured cells by a mechanism that is different from that by which it binds to collagen.

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Selected References

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

  1. Barnes M. J., Morton L. F., Levene C. I. Synthesis of interstitial collagens by pig aortic endothelial cells in culture. Biochem Biophys Res Commun. 1978 Oct 16;84(3):646–653. doi: 10.1016/0006-291x(78)90754-4. [DOI] [PubMed] [Google Scholar]
  2. Baumgartner H. R. The role of blood flow in platelet adhesion, fibrin deposition, and formation of mural thrombi. Microvasc Res. 1973 Mar;5(2):167–179. doi: 10.1016/0026-2862(73)90069-1. [DOI] [PubMed] [Google Scholar]
  3. Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  4. Hormia M., Lehto V. P., Virtanen I. Factor VIII-related antigen. A pericellular matrix component of cultured human endothelial cells. Exp Cell Res. 1983 Dec;149(2):483–497. doi: 10.1016/0014-4827(83)90360-9. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Hovig T., Stormorken H. Ultrastructural studies on the platelet plug formation in bleeding time wounds from normal individuals and patients with von Willebrand's disease. Acta Pathol Microbiol Scand Suppl. 1974;Suppl 248:105–122. [PubMed] [Google Scholar]
  8. Jaffe E. A., Nachman R. L., Becker C. G., Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. doi: 10.1172/JCI107470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Legrand Y. J., Rodriguez-Zeballos A., Kartalis G., Fauvel F., Caen J. P. Adsorption of factor VIII antigen-activity complex by collagen. Thromb Res. 1978 Nov;13(5):909–911. doi: 10.1016/0049-3848(78)90196-2. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Rand J. H., Sussman I. I., Gordon R. E., Chu S. V., Solomon V. Localization of factor-VIII-related antigen in human vascular subendothelium. Blood. 1980 May;55(5):752–756. [PubMed] [Google Scholar]
  13. Ross R., Glomset J. A. The pathogenesis of atherosclerosis (second of two parts). N Engl J Med. 1976 Aug 19;295(8):420–425. doi: 10.1056/NEJM197608192950805. [DOI] [PubMed] [Google Scholar]
  14. Ross R. The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol. 1971 Jul;50(1):172–186. doi: 10.1083/jcb.50.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ruggeri Z. M., Bader R., de Marco L. Glanzmann thrombasthenia: deficient binding of von Willebrand factor to thrombin-stimulated platelets. Proc Natl Acad Sci U S A. 1982 Oct;79(19):6038–6041. doi: 10.1073/pnas.79.19.6038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ruggeri Z. M., De Marco L., Gatti L., Bader R., Montgomery R. R. Platelets have more than one binding site for von Willebrand factor. J Clin Invest. 1983 Jul;72(1):1–12. doi: 10.1172/JCI110946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sage H., Crouch E., Bornstein P. Collagen synthesis by bovine aortic endothelial cells in culture. Biochemistry. 1979 Nov 27;18(24):5433–5442. doi: 10.1021/bi00591a028. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Sakariassen K. S., Bolhuis P. A., Sixma J. J. Human blood platelet adhesion to artery subendothelium is mediated by factor VIII-Von Willebrand factor bound to the subendothelium. Nature. 1979 Jun 14;279(5714):636–638. doi: 10.1038/279636a0. [DOI] [PubMed] [Google Scholar]
  20. Sakariassen K. S., Bolhuis P. A., Sixma J. J. Platelet adherence to subendothelium of human arteries in pulsatile and steady flow. 1980 Aug 15-Sep 1Thromb Res. 19(4-5):547–559. doi: 10.1016/0049-3848(80)90027-4. [DOI] [PubMed] [Google Scholar]
  21. Santoro S. A., Cowan J. F. Adsorption of von Willebrand factor by fibrillar collagen--implications concerning the adhesion of platelets to collagen. Coll Relat Res. 1982 Jan;2(1):31–43. doi: 10.1016/s0174-173x(82)80039-3. [DOI] [PubMed] [Google Scholar]
  22. Santoro S. A. Preferential binding of high molecular weight forms of von Willebrand factor to fibrillar collagen. Biochim Biophys Acta. 1983 Mar 15;756(1):123–126. doi: 10.1016/0304-4165(83)90032-6. [DOI] [PubMed] [Google Scholar]
  23. Scott D. M., Griffin B., Pepper D. S., Barnes M. J. The binding of purified factor VIII/von Willebrand factor to collagens of differing type and form. Thromb Res. 1981 Dec 1;24(5-6):467–472. doi: 10.1016/0049-3848(81)90080-3. [DOI] [PubMed] [Google Scholar]
  24. Sixma J. J., Sakariassen K. S., Stel H. V., Houdijk W. P., In der Maur D. W., Hamer R. J., de Groot P. G., van Mourik J. A. Functional domains on von Willebrand factor. Recognition of discrete tryptic fragments by monoclonal antibodies that inhibit interaction of von Willebrand factor with platelets and with collagen. J Clin Invest. 1984 Sep;74(3):736–744. doi: 10.1172/JCI111489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stel H. V., Sakariassen K. S., Scholte B. J., Veerman E. C., van der Kwast T. H., de Groot P. G., Sixma J. J., van Mourik J. A. Characterization of 25 monoclonal antibodies to factor VIII-von Willebrand factor: relationship between ristocetin-induced platelet aggregation and platelet adherence to subendothelium. Blood. 1984 Jun;63(6):1408–1415. [PubMed] [Google Scholar]
  26. 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]
  27. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tschopp T. B., Weiss H. J., Baumgartner H. R. Decreased adhesion of platelets to subendothelium in von Willebrand's disease. J Lab Clin Med. 1974 Feb;83(2):296–300. [PubMed] [Google Scholar]
  29. Verweij C. L., de Vries C. J., Distel B., van Zonneveld A. J., van Kessel A. G., van Mourik J. A., Pannekoek H. Construction of cDNA coding for human von Willebrand factor using antibody probes for colony-screening and mapping of the chromosomal gene. Nucleic Acids Res. 1985 Jul 11;13(13):4699–4717. doi: 10.1093/nar/13.13.4699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wagner D. D., Marder V. J. Biosynthesis of von Willebrand protein by human endothelial cells. Identification of a large precursor polypeptide chain. J Biol Chem. 1983 Feb 25;258(4):2065–2067. [PubMed] [Google Scholar]
  31. Wagner D. D., Urban-Pickering M., Marder V. J. Von Willebrand protein binds to extracellular matrices independently of collagen. Proc Natl Acad Sci U S A. 1984 Jan;81(2):471–475. doi: 10.1073/pnas.81.2.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. van Mourik J. A., Mochtar I. A. Purification of human antihemophilic factor (factor VIII) by gel chromatography. Biochim Biophys Acta. 1970 Dec 22;221(3):677–679. doi: 10.1016/0005-2795(70)90247-3. [DOI] [PubMed] [Google Scholar]

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