Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1988 May;81(5):1600–1607. doi: 10.1172/JCI113494

Identification and isolation of a platelet GPIb-like protein in human umbilical vein endothelial cells and bovine aortic smooth muscle cells.

A S Asch 1, B Adelman 1, M Fujimoto 1, R L Nachman 1
PMCID: PMC442595  PMID: 3284916

Abstract

Glycoprotein Ib (GPIb) is an intrinsic platelet membrane protein that plays a major role in platelet adherence and mediates ristocetin-dependent platelet von Willebrand factor binding. Recent reports that the platelet membrane glycoprotein complex IIb/IIIa is expressed in several cell types prompted us to look for GPIb expression in other vascular cells. Immunoperoxidase staining of human stomach and skin histologic sections with polyclonal as well as monoclonal anti-GPIb antibody revealed the presence of GPIb in the endothelial cell and smooth muscle cell layers. Western blotting using monospecific polyclonal anti-GPIb antibodies confirmed the presence of immunoreactive GPIb in human umbilical vein endothelial and bovine aortic smooth muscle cell cultures. Fab fragments of a monoclonal anti-GPIb antibody were used to immunoprecipitate [3H]leucine labeled GPIb from metabolically labeled cells. The GPIb in these cells was functional as measured by ristocetin-dependent cell agglutination and by vWF binding. Endothelial cells as well as smooth muscle cells bound 125I-labeled vWF in a ristocetin-dependent manner, with a Kd of 7.9 nM.

Full text

PDF
1600

Images in this article

1602Image
on p.1602
1602Image
on p.1602
1602Image
on p.1602
1602Image
on p.1602
1601Image
on p.1601
1601Image
on p.1601
1601Image
on p.1601
1601Image
on p.1601
1603Image
on p.1603
1604Image
on p.1604
1604Image
on p.1604
1604Image
on p.1604
1604Image
on p.1604
1604Image
on p.1604
1605Image
on p.1605
1605Image
on p.1605
1605Image
on p.1605

Selected References

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

  1. 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]
  2. Becker C. G., Hajjar D. P., Hefton J. M. Tobacco constituents are mitogenic for arterial smooth-muscle cells. Am J Pathol. 1985 Jul;120(1):1–5. [PMC free article] [PubMed] [Google Scholar]
  3. Brown S. S., Malinoff H. L., Wicha M. S. Connectin: cell surface protein that binds both laminin and actin. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5927–5930. doi: 10.1073/pnas.80.19.5927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Casonato A., Fabris F., Vicariotto M., Girolami A. The evaluation of factor VIII antigen by means of a simple slide test. Am J Clin Pathol. 1985 Jul;84(1):107–111. doi: 10.1093/ajcp/84.1.107. [DOI] [PubMed] [Google Scholar]
  5. Charo I. F., Fitzgerald L. A., Steiner B., Rall S. C., Jr, Bekeart L. S., Phillips D. R. Platelet glycoproteins IIb and IIIa: evidence for a family of immunologically and structurally related glycoproteins in mammalian cells. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8351–8355. doi: 10.1073/pnas.83.21.8351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cines D. B., Tomaski A., Tannenbaum S. Immune endothelial-cell injury in heparin-associated thrombocytopenia. N Engl J Med. 1987 Mar 5;316(10):581–589. doi: 10.1056/NEJM198703053161004. [DOI] [PubMed] [Google Scholar]
  7. Duband J. L., Rocher S., Chen W. T., Yamada K. M., Thiery J. P. Cell adhesion and migration in the early vertebrate embryo: location and possible role of the putative fibronectin receptor complex. J Cell Biol. 1986 Jan;102(1):160–178. doi: 10.1083/jcb.102.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fitzgerald L. A., Charo I. F., Phillips D. R. Human and bovine endothelial cells synthesize membrane proteins similar to human platelet glycoproteins IIb and IIIa. J Biol Chem. 1985 Sep 15;260(20):10893–10896. [PubMed] [Google Scholar]
  9. Fox J. E. Linkage of a membrane skeleton to integral membrane glycoproteins in human platelets. Identification of one of the glycoproteins as glycoprotein Ib. J Clin Invest. 1985 Oct;76(4):1673–1683. doi: 10.1172/JCI112153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jaffe E. A., Hoyer L. W., Nachman R. L. Synthesis of von Willebrand factor by cultured human endothelial cells. Proc Natl Acad Sci U S A. 1974 May;71(5):1906–1909. doi: 10.1073/pnas.71.5.1906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Kao K. J., Pizzo S. V., McKee P. A. Demonstration and characterization of specific binding sites for factor VIII/von Willebrand factor on human platelets. J Clin Invest. 1979 Apr;63(4):656–664. doi: 10.1172/JCI109348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kieffer N., Debili N., Wicki A., Titeux M., Henri A., Mishal Z., Breton-Gorius J., Vainchenker W., Clemetson K. J. Expression of platelet glycoprotein Ib alpha in HEL cells. J Biol Chem. 1986 Dec 5;261(34):15854–15862. [PubMed] [Google Scholar]
  14. Köhler G., Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur J Immunol. 1976 Jul;6(7):511–519. doi: 10.1002/eji.1830060713. [DOI] [PubMed] [Google Scholar]
  15. Lopez J. A., Chung D. W., Fujikawa K., Hagen F. S., Papayannopoulou T., Roth G. J. Cloning of the alpha chain of human platelet glycoprotein Ib: a transmembrane protein with homology to leucine-rich alpha 2-glycoprotein. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5615–5619. doi: 10.1073/pnas.84.16.5615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maciag T., Hoover G. A., Stemerman M. B., Weinstein R. Serial propagation of human endothelial cells in vitro. J Cell Biol. 1981 Nov;91(2 Pt 1):420–426. doi: 10.1083/jcb.91.2.420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  18. Michelson A. D., Loscalzo J., Melnick B., Coller B. S., Handin R. I. Partial characterization of a binding site for von Willebrand factor on glycocalicin. Blood. 1986 Jan;67(1):19–26. [PubMed] [Google Scholar]
  19. Montgomery R. R., Kunicki T. J., Taves C., Pidard D., Corcoran M. Diagnosis of Bernard-Soulier syndrome and Glanzmann's thrombasthenia with a monoclonal assay on whole blood. J Clin Invest. 1983 Feb;71(2):385–389. doi: 10.1172/JCI110780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Morisato D. K., Gralnick H. R. Selective binding of the factor VIII/von Willebrand factor protein to human platelets. Blood. 1980 Jan;55(1):9–15. [PubMed] [Google Scholar]
  21. Nachman R. L., Kinoshita T., Ferris B. Structural analysis of human platelet membrane glycoprotein I complex. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2952–2954. doi: 10.1073/pnas.76.6.2952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nurden A. T., Caen J. P. The different glycoprotein abnormalities in thrombasthenic and Bernard-Soulier platelets. Semin Hematol. 1979 Jul;16(3):234–250. [PubMed] [Google Scholar]
  23. Phillips D. R., Agin P. P. Platelet plasma membrane glycoproteins. Evidence for the presence of nonequivalent disulfide bonds using nonreduced-reduced two-dimensional gel electrophoresis. J Biol Chem. 1977 Mar 25;252(6):2121–2126. [PubMed] [Google Scholar]
  24. Solum N. O., Olsen T. M. Effects of diamide and dibucaine on platelet glycoprotein Ib, actin-binding protein and cytoskeleton. Biochim Biophys Acta. 1985 Jul 25;817(2):249–260. doi: 10.1016/0005-2736(85)90026-4. [DOI] [PubMed] [Google Scholar]
  25. Solum N. O., Olsen T. M. Glycoprotein Ib in the Triton-insoluble (cytoskeletal) fraction of blood platelets. Biochim Biophys Acta. 1984 Jun 29;799(3):209–220. doi: 10.1016/0304-4165(84)90263-0. [DOI] [PubMed] [Google Scholar]
  26. Sprandio J. D., Shapiro S. S., Thiagarajan P., McCord S. Cultured human umbilical vein endothelial cells contain a membrane glycoprotein immunologically related to platelet glycoprotein Ib. Blood. 1988 Jan;71(1):234–237. [PubMed] [Google Scholar]
  27. Stricker R. B., Wong D., Saks S. R., Corash L., Shuman M. A. Acquired Bernard-Soulier syndrome. Evidence for the role of a 210,000-molecular weight protein in the interaction of platelets with von Willebrand factor. J Clin Invest. 1985 Sep;76(3):1274–1278. doi: 10.1172/JCI112084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thiagarajan P., Shapiro S. S., Levine E., DeMarco L., Yalcin A. A monoclonal antibody to human platelet glycoprotein IIIa detects a related protein in cultured human endothelial cells. J Clin Invest. 1985 Mar;75(3):896–901. doi: 10.1172/JCI111789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Thornton S. C., Mueller S. N., Levine E. M. Human endothelial cells: use of heparin in cloning and long-term serial cultivation. Science. 1983 Nov 11;222(4624):623–625. doi: 10.1126/science.6635659. [DOI] [PubMed] [Google Scholar]
  30. Titani K., Takio K., Handa M., Ruggeri Z. M. Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5610–5614. doi: 10.1073/pnas.84.16.5610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weiss H. J., Hoyer L. W., Rickles F. R., Varma A., Rogers J. Quantitative assay of a plasma factor deficient in von Willebrand's disease that is necessary for platelet aggregation. Relationship to factor VIII procoagulant activity and antigen content. J Clin Invest. 1973 Nov;52(11):2708–2716. doi: 10.1172/JCI107465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wheeler M. E., Cox A. C., Carroll R. C. Retention of the glycoprotein IIb-IIIa complex in the isolated platelet cytoskeleton. Effects of separable assembly of platelet pseudopodal and contractile cytoskeletons. J Clin Invest. 1984 Sep;74(3):1080–1089. doi: 10.1172/JCI111475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. de Groot P. G., Gonsalves M. D., Loesberg C., van Buul-Wortelboer M. F., van Aken W. G., van Mourik J. A. Thrombin-induced release of von Willebrand factor from endothelial cells is mediated by phospholipid methylation. Prostacyclin synthesis is independent of phospholipid methylation. J Biol Chem. 1984 Nov 10;259(21):13329–13333. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES