Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Dec 1;99(6):2056–2060. doi: 10.1083/jcb.99.6.2056

Platelet-collagen adhesion: inhibition by a monoclonal antibody that binds glycoprotein IIb

PMCID: PMC2113561  PMID: 6238974

Abstract

To identify platelet surface structures involved in adhesion to collagen, the effect of 16 murine antiplatelet membrane hybridoma antibodies were tested in a defined, in vitro assay. Four of these antibodies inhibited platelet-collagen adhesion and reacted with a polypeptide with Mr approximately 125,000, as determined by immunoblots after gel electrophoresis under reducing conditions. Through detailed studies with one of these antibodies, the monoclonal antibody PMI-1, the relevant antigen was identified as platelet glycoprotein IIb alpha, based upon (a) co-migration with this glycoprotein in two-dimensional gel electrophoresis and (b) co-purification by immunoaffinity chromatography with a protein with apparent Mr identical to that of glycoprotein III, under conditions in which glycoproteins IIb and III form a complex. Univalent antibody fragments prepared from monoclonal antibody PMI-1 inhibited greater than 80% of platelet-collagen adhesion, and inhibition was completely blocked by the immunopurified antigen. These results indicate that glycoprotein IIb participates in some aspect of platelet-collagen adhesion. In contrast, the purified antigen only partially neutralized a polyclonal antiserum that blocked platelet-collagen adhesion, to a maximum of approximately 25%, at saturating antigen concentrations. Thus, by these immunological criteria, glycoprotein IIb is not the only molecule involved in this process.

Full Text

The Full Text of this article is available as a PDF (953.2 KB).

Selected References

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

  1. Barber A. J., Jamieson G. A. Isolation and characterization of plasma membranes from human blood platelets. J Biol Chem. 1970 Dec 10;245(23):6357–6365. [PubMed] [Google Scholar]
  2. Bordier C. Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem. 1981 Feb 25;256(4):1604–1607. [PubMed] [Google Scholar]
  3. Chiang T. M., Kang A. H. Isolation and purification of collagen alpha 1(I) receptor from human platelet membrane. J Biol Chem. 1982 Jul 10;257(13):7581–7586. [PubMed] [Google Scholar]
  4. Curtiss L. K., Edgington T. S. Immunochemical heterogeneity of human plasma apolipoprotein B. I. Apolipoprotein B binding of mouse hybridoma antibodies. J Biol Chem. 1982 Dec 25;257(24):15213–15221. [PubMed] [Google Scholar]
  5. Edelman G. M. Cell adhesion molecules. Science. 1983 Feb 4;219(4584):450–457. doi: 10.1126/science.6823544. [DOI] [PubMed] [Google Scholar]
  6. Gefter M. L., Margulies D. H., Scharff M. D. A simple method for polyethylene glycol-promoted hybridization of mouse myeloma cells. Somatic Cell Genet. 1977 Mar;3(2):231–236. doi: 10.1007/BF01551818. [DOI] [PubMed] [Google Scholar]
  7. Kotite N. J., Staros J. V., Cunningham L. W. Interaction of specific platelet membrane proteins with collagen: evidence from chemical cross-linking. Biochemistry. 1984 Jun 19;23(13):3099–3104. doi: 10.1021/bi00308a038. [DOI] [PubMed] [Google Scholar]
  8. Levy-Toledano S., Tobelem G., Legrand C., Bredoux R., Degos L., Nurden A., Caen J. P. Acquired IgG antibody occurring in a thrombasthenic patient: its effect on human platelet function. Blood. 1978 Jun;51(6):1065–1071. [PubMed] [Google Scholar]
  9. McEver R. P., Baenziger N. L., Majerus P. W. Isolation and quantitation of the platelet membrane glycoprotein deficient in thrombasthenia using a monoclonal hybridoma antibody. J Clin Invest. 1980 Dec;66(6):1311–1318. doi: 10.1172/JCI109983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. McEver R. P., Bennett E. M., Martin M. N. Identification of two structurally and functionally distinct sites on human platelet membrane glycoprotein IIb-IIIa using monoclonal antibodies. J Biol Chem. 1983 Apr 25;258(8):5269–5275. [PubMed] [Google Scholar]
  11. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  12. Neff N. T., Lowrey C., Decker C., Tovar A., Damsky C., Buck C., Horwitz A. F. A monoclonal antibody detaches embryonic skeletal muscle from extracellular matrices. J Cell Biol. 1982 Nov;95(2 Pt 1):654–666. doi: 10.1083/jcb.95.2.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nurden A. T., Caen J. P. Role of surface glycoproteins in human platelet function. Thromb Haemost. 1976 Feb 29;35(1):139–150. [PubMed] [Google Scholar]
  14. Nurden A. T., Caen J. P. Specific roles for platelet surface glycoproteins in platelet function. Nature. 1975 Jun 26;255(5511):720–722. doi: 10.1038/255720a0. [DOI] [PubMed] [Google Scholar]
  15. Painter R. G., Ginsberg M. Concanavalin A induces interactions between surface glycoproteins and the platelet cytoskeleton. J Cell Biol. 1982 Feb;92(2):565–573. doi: 10.1083/jcb.92.2.565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Pidard D., Montgomery R. R., Bennett J. S., Kunicki T. J. Interaction of AP-2, a monoclonal antibody specific for the human platelet glycoprotein IIb-IIIa complex, with intact platelets. J Biol Chem. 1983 Oct 25;258(20):12582–12586. [PubMed] [Google Scholar]
  18. Shadle P. J., Barondes S. H. Adhesion of human platelets to immobilized trimeric collagen. J Cell Biol. 1982 Oct;95(1):361–365. doi: 10.1083/jcb.95.1.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shadle P. J., Barondes S. H. Platelet-collagen adhesion: evidence for participation of antigenically distinct entities. J Cell Biol. 1984 Dec;99(6):2048–2055. doi: 10.1083/jcb.99.6.2048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Springer W. R., Barondes S. H. Monoclonal antibodies block cell-cell adhesion in Dictyostelium discoideum. J Biol Chem. 1983 Apr 25;258(8):4698–4701. [PubMed] [Google Scholar]
  21. Tsunehisa S., Tsuji T., Tohyama H., Osawa T. Interaction of human platelet membrane glycoproteins with collagen and lectins. Biochim Biophys Acta. 1984 Jan 24;797(1):10–19. [PubMed] [Google Scholar]
  22. Vollmers H. P., Birchmeier W. Monoclonal antibodies inhibit the adhesion of mouse B 16 melanoma cells in vitro and block lung metastasis in vivo. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3729–3733. doi: 10.1073/pnas.80.12.3729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weiss H. J. Platelet physiology and abnormalities of platelet function (first of two parts). N Engl J Med. 1975 Sep 11;293(11):531–541. doi: 10.1056/NEJM197509112931105. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES