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. 1986 Nov;83(21):8351–8355. doi: 10.1073/pnas.83.21.8351

Platelet glycoproteins IIb and IIIa: evidence for a family of immunologically and structurally related glycoproteins in mammalian cells.

I F Charo, L A Fitzgerald, B Steiner, S C Rall Jr, L S Bekeart, D R Phillips
PMCID: PMC386926  PMID: 3534886

Abstract

Human and bovine cultured cell lines and circulating leukocytes were examined for the presence of surface proteins similar to platelet glycoproteins IIb (GPIIb) and IIIa (GPIIIa). Human endothelial cells, smooth muscle cells, and MG-63 fibroblast-like cells were found to have surface proteins that cross-reacted with platelet GPIIb and GPIIIa antibodies, existed as complexes, and had molecular weights similar to those of the corresponding platelet glycoproteins. Bovine endothelial cells and smooth muscle cells also expressed GPIIb- and GPIIIa-like surface proteins. Metabolic labeling studies with [35S]methionine demonstrated that the cultured cells synthesized these glycoproteins. The GPIIIa-like protein in human endothelial and smooth muscle cells had the same isoelectric point as platelet GPIIIa, whereas their GPIIb alpha-like protein was slightly more acidic than platelet GPIIb alpha (pI = 5.2-5.3 versus 5.5). Platelet and endothelial cell GPIIb alpha (but not GPIIIa) showed an increased electrophoretic mobility in Ca2+ -containing versus EDTA-containing gels, implying a Ca2+ -GPIIb alpha interaction. The amino acid sequence of the amino termini of platelet GPIIb alpha and GPIIb beta and of the alpha chains of the leukocyte LFA-1 and Mac-1 glycoprotein complexes had significant sequence homology. These data indicate that glycoproteins that have either immunological cross-reactivity or amino-terminal sequence homology with the platelet GPIIb-IIIa complex are widely distributed in human and non-human adherent cells and circulating leukocytes and suggest that these proteins may be the products of a large gene family whose expression is cell specific.

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Selected References

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  1. Bennett J. S., Hoxie J. A., Leitman S. F., Vilaire G., Cines D. B. Inhibition of fibrinogen binding to stimulated human platelets by a monoclonal antibody. Proc Natl Acad Sci U S A. 1983 May;80(9):2417–2421. doi: 10.1073/pnas.80.9.2417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brass L. F. Ca2+ homeostasis in unstimulated platelets. J Biol Chem. 1984 Oct 25;259(20):12563–12570. [PubMed] [Google Scholar]
  3. Bray P. F., Rosa J. P., Lingappa V. R., Kan Y. W., McEver R. P., Shuman M. A. Biogenesis of the platelet receptor for fibrinogen: evidence for separate precursors for glycoproteins IIb and IIIa. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1480–1484. doi: 10.1073/pnas.83.5.1480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burgess W. H., Jemiolo D. K., Kretsinger R. H. Interaction of calcium and calmodulin in the presence of sodium dodecyl sulfate. Biochim Biophys Acta. 1980 Jun 26;623(2):257–270. doi: 10.1016/0005-2795(80)90254-8. [DOI] [PubMed] [Google Scholar]
  5. Coller B. S. A new murine monoclonal antibody reports an activation-dependent change in the conformation and/or microenvironment of the platelet glycoprotein IIb/IIIa complex. J Clin Invest. 1985 Jul;76(1):101–108. doi: 10.1172/JCI111931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cosgrove L. J., Sandrin M. S., Rajasekariah P., McKenzie I. F. A genomic clone encoding the alpha chain of the OKM1, LFA-1, and platelet glycoprotein IIb-IIIa molecules. Proc Natl Acad Sci U S A. 1986 Feb;83(3):752–756. doi: 10.1073/pnas.83.3.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cox J. A., Stein E. A. Characterization of a new sarcoplasmic calcium-binding protein with magnesium-induced cooperativity in the binding of calcium. Biochemistry. 1981 Sep 15;20(19):5430–5436. doi: 10.1021/bi00522a012. [DOI] [PubMed] [Google Scholar]
  8. Dana N., Todd R. F., 3rd, Pitt J., Springer T. A., Arnaout M. A. Deficiency of a surface membrane glycoprotein (Mo1) in man. J Clin Invest. 1984 Jan;73(1):153–159. doi: 10.1172/JCI111186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Edelman G. M. Cell adhesion and the molecular processes of morphogenesis. Annu Rev Biochem. 1985;54:135–169. doi: 10.1146/annurev.bi.54.070185.001031. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Fitzgerald L. A., Leung B., Phillips D. R. A method for purifying the platelet membrane glycoprotein IIb-IIIa complex. Anal Biochem. 1985 Nov 15;151(1):169–177. doi: 10.1016/0003-2697(85)90067-3. [DOI] [PubMed] [Google Scholar]
  12. Fitzgerald L. A., Phillips D. R. Calcium regulation of the platelet membrane glycoprotein IIb-IIIa complex. J Biol Chem. 1985 Sep 15;260(20):11366–11374. [PubMed] [Google Scholar]
  13. Gimbrone M. A., Jr, Cotran R. S. Human vascular smooth muscle in culture. Growth and ultrastructure. Lab Invest. 1975 Jul;33(1):16–27. [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  17. Phillips D. R., Agin P. P. Platelet membrane defects in Glanzmann's thrombasthenia. Evidence for decreased amounts of two major glycoproteins. J Clin Invest. 1977 Sep;60(3):535–545. doi: 10.1172/JCI108805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Phillips D. R. Effect of trypsin on the exposed polypeptides and glycoproteins in the human platelet membrane. Biochemistry. 1972 Nov 21;11(24):4582–4588. doi: 10.1021/bi00774a025. [DOI] [PubMed] [Google Scholar]
  19. Plow E. F., Ginsberg M. H. Specific and saturable binding of plasma fibronectin to thrombin-stimulated human platelets. J Biol Chem. 1981 Sep 25;256(18):9477–9482. [PubMed] [Google Scholar]
  20. Rougon G., Marshak D. R. Structural and immunological characterization of the amino-terminal domain of mammalian neural cell adhesion molecules. J Biol Chem. 1986 Mar 5;261(7):3396–3401. [PubMed] [Google Scholar]
  21. 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]
  22. Sanchez-Madrid F., Nagy J. A., Robbins E., Simon P., Springer T. A. A human leukocyte differentiation antigen family with distinct alpha-subunits and a common beta-subunit: the lymphocyte function-associated antigen (LFA-1), the C3bi complement receptor (OKM1/Mac-1), and the p150,95 molecule. J Exp Med. 1983 Dec 1;158(6):1785–1803. doi: 10.1084/jem.158.6.1785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Springer T. A., Teplow D. B., Dreyer W. J. Sequence homology of the LFA-1 and Mac-1 leukocyte adhesion glycoproteins and unexpected relation to leukocyte interferon. Nature. 1985 Apr 11;314(6011):540–542. doi: 10.1038/314540a0. [DOI] [PubMed] [Google Scholar]
  24. Stephens R. E. High-resolution preparative SDS-polyacrylamide gel electrophoresis: fluorescent visualization and electrophoretic elution-concentration of protein bands. Anal Biochem. 1975 May 12;65(1-2):369–379. doi: 10.1016/0003-2697(75)90521-7. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Weisgraber K. H., Rall S. C., Jr, Mahley R. W. Human E apoprotein heterogeneity. Cysteine-arginine interchanges in the amino acid sequence of the apo-E isoforms. J Biol Chem. 1981 Sep 10;256(17):9077–9083. [PubMed] [Google Scholar]

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