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. 1990 Nov;1(12):883–893. doi: 10.1091/mbc.1.12.883

Affinity modulation of the alpha IIb beta 3 integrin (platelet GPIIb-IIIa) is an intrinsic property of the receptor.

T E O'Toole 1, J C Loftus 1, X P Du 1, A A Glass 1, Z M Ruggeri 1, S J Shattil 1, E F Plow 1, M H Ginsberg 1
PMCID: PMC362859  PMID: 2100193

Abstract

To analyze the basis of affinity modulation of integrin function, we studied cloned stable Chinese hamster ovary cell lines expressing recombinant integrins of the beta 3 family (alpha IIb beta 3 and alpha v beta 3). Antigenic and peptide recognition specificities of the recombinant receptors resembled those of the native receptors found in platelets or endothelial cells. The alpha IIb beta 3-expressing cell line (A5) bound RGD peptides and immobilized fibrinogen (Fg) but not soluble fibrinogen or the activation-specific monoclonal anti-alpha IIb beta 3 (PAC1), indicating that it was in the affinity state found on resting platelets. Several platelet agonists failed to alter the affinity state of ("activate") recombinant alpha IIb beta 3. The binding of soluble Fg and PAC1, however, was stimulated in both platelets and A5 cells by addition of IgG papain-digestion products (Fab) fragments of certain beta 3-specific monoclonal antibodies. These antibodies stimulated PAC1 binding to platelets fixed under conditions rendering them unresponsive to other agonists. Addition of these antibodies to detergent-solubilized alpha IIb beta 3 also stimulated specific Fg binding. These data demonstrate that certain anti-beta 3 antibodies activate alpha IIb beta 3 by acting directly on the receptor, possibly by altering its conformation. Furthermore, they indicate that the activation state of alpha IIb beta 3 is a property of the receptor itself rather than of the surrounding cell membrane microenvironment.

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Selected References

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

  1. Altieri D. C., Bader R., Mannucci P. M., Edgington T. S. Oligospecificity of the cellular adhesion receptor Mac-1 encompasses an inducible recognition specificity for fibrinogen. J Cell Biol. 1988 Nov;107(5):1893–1900. doi: 10.1083/jcb.107.5.1893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bennett J. S., Vilaire G. Exposure of platelet fibrinogen receptors by ADP and epinephrine. J Clin Invest. 1979 Nov;64(5):1393–1401. doi: 10.1172/JCI109597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bodary S. C., Napier M. A., McLean J. W. Expression of recombinant platelet glycoprotein IIbIIIa results in a functional fibrinogen-binding complex. J Biol Chem. 1989 Nov 15;264(32):18859–18862. [PubMed] [Google Scholar]
  4. Boucheix C., Soria C., Mirshahi M., Soria J., Perrot J. Y., Fournier N., Billard M., Rosenfeld C. Characteristics of platelet aggregation induced by the monoclonal antibody ALB6 (acute lymphoblastic leukemia antigen p 24). Inhibition of aggregation by ALB6Fab. FEBS Lett. 1983 Sep 19;161(2):289–295. doi: 10.1016/0014-5793(83)81027-8. [DOI] [PubMed] [Google Scholar]
  5. Cheresh D. A., Berliner S. A., Vicente V., Ruggeri Z. M. Recognition of distinct adhesive sites on fibrinogen by related integrins on platelets and endothelial cells. Cell. 1989 Sep 8;58(5):945–953. doi: 10.1016/0092-8674(89)90946-x. [DOI] [PubMed] [Google Scholar]
  6. Coller B. S. Activation affects access to the platelet receptor for adhesive glycoproteins. J Cell Biol. 1986 Aug;103(2):451–456. doi: 10.1083/jcb.103.2.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coller B. S. Interaction of normal, thrombasthenic, and Bernard-Soulier platelets with immobilized fibrinogen: defective platelet-fibrinogen interaction in thrombasthenia. Blood. 1980 Feb;55(2):169–178. [PubMed] [Google Scholar]
  8. D'Souza S. E., Ginsberg M. H., Burke T. A., Plow E. F. The ligand binding site of the platelet integrin receptor GPIIb-IIIa is proximal to the second calcium binding domain of its alpha subunit. J Biol Chem. 1990 Feb 25;265(6):3440–3446. [PubMed] [Google Scholar]
  9. Detmers P. A., Wright S. D., Olsen E., Kimball B., Cohn Z. A. Aggregation of complement receptors on human neutrophils in the absence of ligand. J Cell Biol. 1987 Sep;105(3):1137–1145. doi: 10.1083/jcb.105.3.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Frelinger A. L., 3rd, Cohen I., Plow E. F., Smith M. A., Roberts J., Lam S. C., Ginsberg M. H. Selective inhibition of integrin function by antibodies specific for ligand-occupied receptor conformers. J Biol Chem. 1990 Apr 15;265(11):6346–6352. [PubMed] [Google Scholar]
  11. Frelinger A. L., 3rd, Lam S. C., Plow E. F., Smith M. A., Loftus J. C., Ginsberg M. H. Occupancy of an adhesive glycoprotein receptor modulates expression of an antigenic site involved in cell adhesion. J Biol Chem. 1988 Sep 5;263(25):12397–12402. [PubMed] [Google Scholar]
  12. Ginsberg M. H., Loftus J. C., Plow E. F. Cytoadhesins, integrins, and platelets. Thromb Haemost. 1988 Feb 25;59(1):1–6. [PubMed] [Google Scholar]
  13. Ginsberg M. H., Loftus J., Ryckwaert J. J., Pierschbacher M., Pytela R., Ruoslahti E., Plow E. F. Immunochemical and amino-terminal sequence comparison of two cytoadhesins indicates they contain similar or identical beta subunits and distinct alpha subunits. J Biol Chem. 1987 Apr 25;262(12):5437–5440. [PubMed] [Google Scholar]
  14. Ginsberg M. H., Taylor L., Painter R. G. The mechanism of thrombin-induced platelet factor 4 secretion. Blood. 1980 Apr;55(4):661–668. [PubMed] [Google Scholar]
  15. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  16. Gulino D., Ryckewaert J. J., Andrieux A., Rabiet M. J., Marguerie G. Identification of a monoclonal antibody against platelet GPIIb that interacts with a calcium-binding site and induces aggregation. J Biol Chem. 1990 Jun 5;265(16):9575–9581. [PubMed] [Google Scholar]
  17. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  18. Jennings L. K., Fox C. F., Kouns W. C., McKay C. P., Ballou L. R., Schultz H. E. The activation of human platelets mediated by anti-human platelet p24/CD9 monoclonal antibodies. J Biol Chem. 1990 Mar 5;265(7):3815–3822. [PubMed] [Google Scholar]
  19. Keizer G. D., Visser W., Vliem M., Figdor C. G. A monoclonal antibody (NKI-L16) directed against a unique epitope on the alpha-chain of human leukocyte function-associated antigen 1 induces homotypic cell-cell interactions. J Immunol. 1988 Mar 1;140(5):1393–1400. [PubMed] [Google Scholar]
  20. Lam S. C., Plow E. F., Smith M. A., Andrieux A., Ryckwaert J. J., Marguerie G., Ginsberg M. H. Evidence that arginyl-glycyl-aspartate peptides and fibrinogen gamma chain peptides share a common binding site on platelets. J Biol Chem. 1987 Jan 25;262(3):947–950. [PubMed] [Google Scholar]
  21. Larson R. S., Hibbs M. L., Springer T. A. The leukocyte integrin LFA-1 reconstituted by cDNA transfection in a nonhematopoietic cell line is functionally active and not transiently regulated. Cell Regul. 1990 Mar;1(4):359–367. doi: 10.1091/mbc.1.4.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Loftus J. C., Albrecht R. M. Redistribution of the fibrinogen receptor of human platelets after surface activation. J Cell Biol. 1984 Sep;99(3):822–829. doi: 10.1083/jcb.99.3.822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lombardo V. T., Hodson E., Roberts J. R., Kunicki T. J., Zimmerman T. S., Ruggeri Z. M. Independent modulation of von Willebrand factor and fibrinogen binding to the platelet membrane glycoprotein IIb/IIIa complex as demonstrated by monoclonal antibody. J Clin Invest. 1985 Nov;76(5):1950–1958. doi: 10.1172/JCI112193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Marguerie G. A., Edgington T. S., Plow E. F. Interaction of fibrinogen with its platelet receptor as part of a multistep reaction in ADP-induced platelet aggregation. J Biol Chem. 1980 Jan 10;255(1):154–161. [PubMed] [Google Scholar]
  25. Marguerie G. A., Plow E. F., Edgington T. S. Human platelets possess an inducible and saturable receptor specific for fibrinogen. J Biol Chem. 1979 Jun 25;254(12):5357–5363. [PubMed] [Google Scholar]
  26. O'Toole T. E., Loftus J. C., Plow E. F., Glass A. A., Harper J. R., Ginsberg M. H. Efficient surface expression of platelet GPIIb-IIIa requires both subunits. Blood. 1989 Jul;74(1):14–18. [PubMed] [Google Scholar]
  27. Parise L. V., Helgerson S. L., Steiner B., Nannizzi L., Phillips D. R. Synthetic peptides derived from fibrinogen and fibronectin change the conformation of purified platelet glycoprotein IIb-IIIa. J Biol Chem. 1987 Sep 15;262(26):12597–12602. [PubMed] [Google Scholar]
  28. Peerschke E. I., Zucker M. B. Fibrinogen receptor exposure and aggregation of human blood platelets produced by ADP and chilling. Blood. 1981 Apr;57(4):663–670. [PubMed] [Google Scholar]
  29. Philips M. R., Buyon J. P., Winchester R., Weissmann G., Abramson S. B. Up-regulation of the iC3b receptor (CR3) is neither necessary nor sufficient to promote neutrophil aggregation. J Clin Invest. 1988 Aug;82(2):495–501. doi: 10.1172/JCI113623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Plow E. F., Loftus J. C., Levin E. G., Fair D. S., Dixon D., Forsyth J., Ginsberg M. H. Immunologic relationship between platelet membrane glycoprotein GPIIb/IIIa and cell surface molecules expressed by a variety of cells. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6002–6006. doi: 10.1073/pnas.83.16.6002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Ruoslahti E., Pierschbacher M. D. New perspectives in cell adhesion: RGD and integrins. Science. 1987 Oct 23;238(4826):491–497. doi: 10.1126/science.2821619. [DOI] [PubMed] [Google Scholar]
  33. Shattil S. J., Brass L. F. Induction of the fibrinogen receptor on human platelets by intracellular mediators. J Biol Chem. 1987 Jan 25;262(3):992–1000. [PubMed] [Google Scholar]
  34. Shattil S. J., Hoxie J. A., Cunningham M., Brass L. F. Changes in the platelet membrane glycoprotein IIb.IIIa complex during platelet activation. J Biol Chem. 1985 Sep 15;260(20):11107–11114. [PubMed] [Google Scholar]
  35. Taub R., Gould R. J., Garsky V. M., Ciccarone T. M., Hoxie J., Friedman P. A., Shattil S. J. A monoclonal antibody against the platelet fibrinogen receptor contains a sequence that mimics a receptor recognition domain in fibrinogen. J Biol Chem. 1989 Jan 5;264(1):259–265. [PubMed] [Google Scholar]
  36. Worthington R. E., Carroll R. C., Boucheix C. Platelet activation by CD9 monoclonal antibodies is mediated by the Fc gamma II receptor. Br J Haematol. 1990 Feb;74(2):216–222. doi: 10.1111/j.1365-2141.1990.tb02568.x. [DOI] [PubMed] [Google Scholar]
  37. Wright S. D., Meyer B. C. Phorbol esters cause sequential activation and deactivation of complement receptors on polymorphonuclear leukocytes. J Immunol. 1986 Mar 1;136(5):1759–1764. [PubMed] [Google Scholar]

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