Skip to main content
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1993 Mar;142(3):843–850.

Co-localization of von Willebrand factor and type VI collagen in human vascular subendothelium.

J H Rand 1, X X Wu 1, B J Potter 1, R R Uson 1, R E Gordon 1
PMCID: PMC1886814  PMID: 8456944

Abstract

The binding of von Willebrand factor (vWF) to subendothelium constitutes an important initial step in the process of platelet adhesion to exposed subendothelium following blood vessel injury. We previously demonstrated that vWF is present in human vascular subendothelium and recently found that a 150 kd vWF-binding protein, which we extracted from subendothelium, is type VI collagen. Although we have established that vWF and type VI collagen bind in vitro, it is not known whether these two proteins are associated in the vascular subendothelium in situ. We, therefore, 1) investigated the morphological effects of our biochemical extraction procedure on human umbilical veins by scanning and transmission electron microscopy, 2) analyzed the subendothelial extract by immunofluorescence for the presence of vWF and collagens and by electron microscopy for morphological characteristics, and 3) localized vWF and type VI collagen in subendothelium by immunofluorescence and by single- and double-label immunoelectron microscopic studies with protein A-conjugated gold particles. We found that the surface exposed following de-endothelialization is composed of microfibrils and contains very little fibrillar collagen. The subendothelium is stripped after sodium dodecyl sulfate-urea extraction, and the extract itself contains immunoreactive vWF and type VI collagen but no immunoreactive type I or III fibrillar collagens. Immunofluorescence and immunoelectron microscopic studies showed that vWF and type VI collagen are both present in subendothelium, where both co-localized to microfibrils. In conclusion, vWF that binds to type VI collagen in vitro, also co-localizes with type VI collagen in subendothelium, where both are associated with microfibrils. Type VI collagen, therefore appears to serve as a biologically significant binding site for vWF in vivo and may thereby play a role in mediating platelet adhesion to exposed subendothelium following vascular injury.

Full text

PDF
843

Images in this article

Selected References

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

  1. Bockenstedt P., McDonagh J., Handin R. I. Binding and covalent cross-linking of purified von Willebrand factor to native monomeric collagen. J Clin Invest. 1986 Aug;78(2):551–556. doi: 10.1172/JCI112608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Colombatti A., Bonaldo P. The superfamily of proteins with von Willebrand factor type A-like domains: one theme common to components of extracellular matrix, hemostasis, cellular adhesion, and defense mechanisms. Blood. 1991 Jun 1;77(11):2305–2315. [PubMed] [Google Scholar]
  3. Fauvel-Lafeve F., Legrand Y. J. Immunochemical identification of a thrombospondin-like structure in an arterial microfibrillar extract. Thromb Res. 1988 Apr 15;50(2):305–316. doi: 10.1016/0049-3848(88)90231-9. [DOI] [PubMed] [Google Scholar]
  4. Fauvel-Lafève F., Legrand Y. J. Binding of plasma von Willebrand factor by arterial microfibrils. Thromb Haemost. 1990 Aug 13;64(1):145–149. [PubMed] [Google Scholar]
  5. Fauvel F., Grant M. E., Legrand Y. J., Souchon H., Tobelem G., Jackson D. S., Caen J. P. Interaction of blood platelets with a microfibrillar extract from adult bovine aorta: requirement for von Willebrand factor. Proc Natl Acad Sci U S A. 1983 Jan;80(2):551–554. doi: 10.1073/pnas.80.2.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Katsuda S., Okada Y., Minamoto T., Oda Y., Matsui Y., Nakanishi I. Collagens in human atherosclerosis. Immunohistochemical analysis using collagen type-specific antibodies. Arterioscler Thromb. 1992 Apr;12(4):494–502. doi: 10.1161/01.atv.12.4.494. [DOI] [PubMed] [Google Scholar]
  7. Kuo H. J., Keene D. R., Glanville R. W. Orientation of type VI collagen monomers in molecular aggregates. Biochemistry. 1989 May 2;28(9):3757–3762. doi: 10.1021/bi00435a020. [DOI] [PubMed] [Google Scholar]
  8. Legrand Y. J., Fauvel F., Arbeille B., Leger D., Mouhli H., Gutman N., Muh J. P. Activation of platelets by microfibrils and collagen. A comparative study. Lab Invest. 1986 May;54(5):566–573. [PubMed] [Google Scholar]
  9. Madri J. A., Dreyer B., Pitlick F. A., Furthmayr H. The collagenous components of the subendothelium. Correlation of structure and function. Lab Invest. 1980 Oct;43(4):303–315. [PubMed] [Google Scholar]
  10. Nyman D. Interaction of collagen with the factor VIII antigen-activity - von Willebrand factor complex. Thromb Res. 1977 Sep;11(3):433–438. doi: 10.1016/0049-3848(77)90196-7. [DOI] [PubMed] [Google Scholar]
  11. Pareti F. I., Niiya K., McPherson J. M., Ruggeri Z. M. Isolation and characterization of two domains of human von Willebrand factor that interact with fibrillar collagen types I and III. J Biol Chem. 1987 Oct 5;262(28):13835–13841. [PubMed] [Google Scholar]
  12. Rand J. H., Gordon R. E., Sussman I. I., Chu S. V., Solomon V. Electron microscopic localization of factor-VIII-related antigen in adult human blood vessels. Blood. 1982 Sep;60(3):627–634. [PubMed] [Google Scholar]
  13. Rand J. H., Patel N. D., Schwartz E., Zhou S. L., Potter B. J. 150-kD von Willebrand factor binding protein extracted from human vascular subendothelium is type VI collagen. J Clin Invest. 1991 Jul;88(1):253–259. doi: 10.1172/JCI115285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Roth G. J. Developing relationships: arterial platelet adhesion, glycoprotein Ib, and leucine-rich glycoproteins. Blood. 1991 Jan 1;77(1):5–19. [PubMed] [Google Scholar]
  16. Roth G. J., Titani K., Hoyer L. W., Hickey M. J. Localization of binding sites within human von Willebrand factor for monomeric type III collagen. Biochemistry. 1986 Dec 30;25(26):8357–8361. doi: 10.1021/bi00374a004. [DOI] [PubMed] [Google Scholar]
  17. Roth J. The preparation of protein A-gold complexes with 3 nm and 15nm gold particles and their use in labelling multiple antigens on ultra-thin sections. Histochem J. 1982 Sep;14(5):791–801. doi: 10.1007/BF01033628. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Ruggeri Z. M., Ware J. The structure and function of von Willebrand factor. Thromb Haemost. 1992 Jun 1;67(6):594–599. [PubMed] [Google Scholar]
  20. 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]
  21. Santoro S. A. Adsorption of von Willebrand factor/factor VIII by the genetically distinct interstitial collagens. Thromb Res. 1981 Mar 15;21(6):689–691. doi: 10.1016/0049-3848(81)90272-3. [DOI] [PubMed] [Google Scholar]
  22. Shekhonin B. V., Domogatsky S. P., Muzykantov V. R., Idelson G. L., Rukosuev V. S. Distribution of type I, III, IV and V collagen in normal and atherosclerotic human arterial wall: immunomorphological characteristics. Coll Relat Res. 1985 Sep;5(4):355–368. doi: 10.1016/s0174-173x(85)80024-8. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Stemerman M. B., Baumgartner H. R., Spaet T. H. The subendothelial microfibril and platelet adhesion. Lab Invest. 1971 Mar;24(3):179–186. [PubMed] [Google Scholar]
  25. Sussman I. I., Rand J. H. Subendothelial deposition of von Willebrand's factor requires the presence of endothelial cells. J Lab Clin Med. 1982 Oct;100(4):526–532. [PubMed] [Google Scholar]
  26. Turitto V. T., Weiss H. J., Baumgartner H. R. Platelet interaction with rabbit subendothelium in von Willebrand's disease: altered thrombus formation distinct from defective platelet adhesion. J Clin Invest. 1984 Nov;74(5):1730–1741. doi: 10.1172/JCI111591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Turitto V. T., Weiss H. J., Zimmerman T. S., Sussman I. I. Factor VIII/von Willebrand factor in subendothelium mediates platelet adhesion. Blood. 1985 Apr;65(4):823–831. [PubMed] [Google Scholar]
  28. Voss B., Rauterberg J. Localization of collagen types I, III, IV and V, fibronectin and laminin in human arteries by the indirect immunofluorescence method. Pathol Res Pract. 1986 Oct;181(5):568–575. doi: 10.1016/S0344-0338(86)80151-0. [DOI] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES