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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1993 Sep 15;90(18):8327–8331. doi: 10.1073/pnas.90.18.8327

Human platelet glycoprotein V: characterization of the polypeptide and the related Ib-V-IX receptor system of adhesive, leucine-rich glycoproteins.

M J Hickey 1, F S Hagen 1, M Yagi 1, G J Roth 1
PMCID: PMC47349  PMID: 7690959

Abstract

Human platelet glycoprotein (GP) V (M(r) 83,300), whose primary structure is reported here, is a part of the Ib-V-IX system of surface glycoproteins (GPs Ib alpha, Ib beta, V, IX) that constitute the receptor for von Willebrand factor (vWf) and mediate the adhesion of platelets to injured vascular surfaces in the arterial circulation, a critical initiating event in hemostasis. System members share physical associations, leucine-rich glycoprotein (LRG) structures, and a congenital deficiency state, Bernard-Soulier syndrome. With PCR techniques and platelet cDNA templates, 1.4 kb of GP V cDNA sequence was obtained that encodes 469 GP V amino acids. A genomic 3.5-kb BamHI fragment was then isolated that includes 3.46 kb of GP V cDNA sequence: the 1.7-kb open reading frame plus 2 bases of the 5' and 1.8 kb of the 3' untranslated regions. Northern blot analysis reveals three GP V platelet transcripts of 3.8, 4.2, and 5.2 kb. A 16-amino acid signal peptide is present. Mature GP V is a 544-amino acid transmembrane protein with a 504-amino acid extracellular domain that encompasses a set of 15 tandem LRG repeats in a "flank-LRG center-flank" array [Roth, G. J. (1991) Blood 77, 5-19] along with eight putative N-linked glycosylation sites and cleavage sites for thrombin and calpain. GP V is a transmembrane, adhesive LRG protein that plays an undefined, but potentially critical, role in the expression and/or function of the Ib-V-IX receptor for vWf/shear-dependent platelet adhesion in arteries.

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  1. Andrews R. K., Fox J. E. Identification of a region in the cytoplasmic domain of the platelet membrane glycoprotein Ib-IX complex that binds to purified actin-binding protein. J Biol Chem. 1992 Sep 15;267(26):18605–18611. [PubMed] [Google Scholar]
  2. Berndt M. C., Gregory C., Kabral A., Zola H., Fournier D., Castaldi P. A. Purification and preliminary characterization of the glycoprotein Ib complex in the human platelet membrane. Eur J Biochem. 1985 Sep 16;151(3):637–649. doi: 10.1111/j.1432-1033.1985.tb09152.x. [DOI] [PubMed] [Google Scholar]
  3. Berndt M. C., Phillips D. R. Purification and preliminary physicochemical characterization of human platelet membrane glycoprotein V. J Biol Chem. 1981 Jan 10;256(1):59–65. [PubMed] [Google Scholar]
  4. Bienz D., Schnippering W., Clemetson K. J. Glycoprotein V is not the thrombin activation receptor on human blood platelets. Blood. 1986 Sep;68(3):720–725. [PubMed] [Google Scholar]
  5. Blochberger T. C., Vergnes J. P., Hempel J., Hassell J. R. cDNA to chick lumican (corneal keratan sulfate proteoglycan) reveals homology to the small interstitial proteoglycan gene family and expression in muscle and intestine. J Biol Chem. 1992 Jan 5;267(1):347–352. [PubMed] [Google Scholar]
  6. Breton-Gorius J., Vainchenker W. Expression of platelet proteins during the in vitro and in vivo differentiation of megakaryocytes and morphological aspects of their maturation. Semin Hematol. 1986 Jan;23(1):43–67. [PubMed] [Google Scholar]
  7. Clemetson K. J., McGregor J. L., James E., Dechavanne M., Lüscher E. F. Characterization of the platelet membrane glycoprotein abnormalities in Bernard-Soulier syndrome and comparison with normal by surface-labeling techniques and high-resolution two-dimensional gel electrophoresis. J Clin Invest. 1982 Aug;70(2):304–311. doi: 10.1172/JCI110618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Greenberg S. M., Rosenthal D. S., Greeley T. A., Tantravahi R., Handin R. I. Characterization of a new megakaryocytic cell line: the Dami cell. Blood. 1988 Dec;72(6):1968–1977. [PubMed] [Google Scholar]
  9. Hickey M. J., Deaven L. L., Roth G. J. Human platelet glycoprotein IX. Characterization of cDNA and localization of the gene to chromosome 3. FEBS Lett. 1990 Nov 12;274(1-2):189–192. doi: 10.1016/0014-5793(90)81361-q. [DOI] [PubMed] [Google Scholar]
  10. Hickey M. J., Roth G. J. Characterization of the gene encoding human platelet glycoprotein IX. J Biol Chem. 1993 Feb 15;268(5):3438–3443. [PubMed] [Google Scholar]
  11. Hickey M. J., Williams S. A., Roth G. J. Human platelet glycoprotein IX: an adhesive prototype of leucine-rich glycoproteins with flank-center-flank structures. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6773–6777. doi: 10.1073/pnas.86.17.6773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kuijper J. L., Wiren K. M., Mathies L. D., Gray C. L., Hagen F. S. Functional cloning vectors for use in directional cDNA cloning using cohesive ends produced with T4 DNA polymerase. Gene. 1992 Mar 15;112(2):147–155. doi: 10.1016/0378-1119(92)90370-5. [DOI] [PubMed] [Google Scholar]
  13. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  14. Long M. W., Heffner C. H., Williams J. L., Peters C., Prochownik E. V. Regulation of megakaryocyte phenotype in human erythroleukemia cells. J Clin Invest. 1990 Apr;85(4):1072–1084. doi: 10.1172/JCI114538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lopez J. A., Chung D. W., Fujikawa K., Hagen F. S., Davie E. W., Roth G. J. The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2135–2139. doi: 10.1073/pnas.85.7.2135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. López J. A., Leung B., Reynolds C. C., Li C. Q., Fox J. E. Efficient plasma membrane expression of a functional platelet glycoprotein Ib-IX complex requires the presence of its three subunits. J Biol Chem. 1992 Jun 25;267(18):12851–12859. [PubMed] [Google Scholar]
  18. López J. A., Ludwig E. H., McCarthy B. J. Polymorphism of human glycoprotein Ib alpha results from a variable number of tandem repeats of a 13-amino acid sequence in the mucin-like macroglycopeptide region. Structure/function implications. J Biol Chem. 1992 May 15;267(14):10055–10061. [PubMed] [Google Scholar]
  19. Martin-Zanca D., Oskam R., Mitra G., Copeland T., Barbacid M. Molecular and biochemical characterization of the human trk proto-oncogene. Mol Cell Biol. 1989 Jan;9(1):24–33. doi: 10.1128/mcb.9.1.24. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Martin P., Papayannopoulou T. HEL cells: a new human erythroleukemia cell line with spontaneous and induced globin expression. Science. 1982 Jun 11;216(4551):1233–1235. doi: 10.1126/science.6177045. [DOI] [PubMed] [Google Scholar]
  21. Mikol D. D., Alexakos M. J., Bayley C. A., Lemons R. S., Le Beau M. M., Stefansson K. Structure and chromosomal localization of the gene for the oligodendrocyte-myelin glycoprotein. J Cell Biol. 1990 Dec;111(6 Pt 1):2673–2679. doi: 10.1083/jcb.111.6.2673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Modderman P. W., Admiraal L. G., Sonnenberg A., von dem Borne A. E. Glycoproteins V and Ib-IX form a noncovalent complex in the platelet membrane. J Biol Chem. 1992 Jan 5;267(1):364–369. [PubMed] [Google Scholar]
  23. Mosher D. F., Vaheri A., Choate J. J., Gahmberg C. G. Action of thrombin on surface glycoproteins of human platelets. Blood. 1979 Mar;53(3):437–445. [PubMed] [Google Scholar]
  24. 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]
  25. Okumura I., Lombart C., Jamieson G. A. Platelet glycocalicin. II. Purification and characterization. J Biol Chem. 1976 Oct 10;251(19):5950–5955. [PubMed] [Google Scholar]
  26. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Phillips D. R., Agin P. P. Platelet plasma membrane glycoproteins. Identification of a proteolytic substrate for thrombin. Biochem Biophys Res Commun. 1977 Apr 25;75(4):940–947. doi: 10.1016/0006-291x(77)91473-5. [DOI] [PubMed] [Google Scholar]
  29. Roth G. J., Church T. A., McMullen B. A., Williams S. A. Human platelet glycoprotein V: a surface leucine-rich glycoprotein related to adhesion. Biochem Biophys Res Commun. 1990 Jul 16;170(1):153–161. doi: 10.1016/0006-291x(90)91253-o. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Roth G. J., Hickey M. J., Chung D. W., Hickstein D. D. Circulating human blood platelets retain appreciable amounts of poly (A)+ RNA. Biochem Biophys Res Commun. 1989 Apr 28;160(2):705–710. doi: 10.1016/0006-291x(89)92490-x. [DOI] [PubMed] [Google Scholar]
  32. Rydel T. J., Ravichandran K. G., Tulinsky A., Bode W., Huber R., Roitsch C., Fenton J. W., 2nd The structure of a complex of recombinant hirudin and human alpha-thrombin. Science. 1990 Jul 20;249(4966):277–280. doi: 10.1126/science.2374926. [DOI] [PubMed] [Google Scholar]
  33. Shimomura T., Fujimura K., Maehama S., Takemoto M., Oda K., Fujimoto T., Oyama R., Suzuki M., Ichihara-Tanaka K., Titani K. Rapid purification and characterization of human platelet glycoprotein V: the amino acid sequence contains leucine-rich repetitive modules as in glycoprotein Ib. Blood. 1990 Jun 15;75(12):2349–2356. [PubMed] [Google Scholar]
  34. Sixma J. J., Wester J. The hemostatic plug. Semin Hematol. 1977 Jul;14(3):265–299. [PubMed] [Google Scholar]
  35. Tschopp T. B., Weiss H. J., Baumgartner H. R. Decreased adhesion of platelets to subendothelium in von Willebrand's disease. J Lab Clin Med. 1974 Feb;83(2):296–300. [PubMed] [Google Scholar]
  36. Vicente V., Houghten R. A., Ruggeri Z. M. Identification of a site in the alpha chain of platelet glycoprotein Ib that participates in von Willebrand factor binding. J Biol Chem. 1990 Jan 5;265(1):274–280. [PubMed] [Google Scholar]
  37. Vu T. K., Hung D. T., Wheaton V. I., Coughlin S. R. Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell. 1991 Mar 22;64(6):1057–1068. doi: 10.1016/0092-8674(91)90261-v. [DOI] [PubMed] [Google Scholar]
  38. Wardell M. R., Reynolds C. C., Berndt M. C., Wallace R. W., Fox J. E. Platelet glycoprotein Ib beta is phosphorylated on serine 166 by cyclic AMP-dependent protein kinase. J Biol Chem. 1989 Sep 15;264(26):15656–15661. [PubMed] [Google Scholar]
  39. Weiss H. J., Tschopp T. B., Baumgartner H. R., Sussman I. I., Johnson M. M., Egan J. J. Decreased adhesion of giant (Bernard-Soulier) platelets to subendothelium. Further implications on the role of the von Willebrand factor in hemostasis. Am J Med. 1974 Dec;57(6):920–925. doi: 10.1016/0002-9343(74)90170-3. [DOI] [PubMed] [Google Scholar]
  40. Wenger R. H., Wicki A. N., Kieffer N., Adolph S., Hameister H., Clemetson K. J. The 5' flanking region and chromosomal localization of the gene encoding human platelet membrane glycoprotein Ib alpha. Gene. 1989 Dec 28;85(2):517–524. doi: 10.1016/0378-1119(89)90446-0. [DOI] [PubMed] [Google Scholar]
  41. Wicki A. N., Clemetson K. J. Structure and function of platelet membrane glycoproteins Ib and V. Effects of leukocyte elastase and other proteases on platelets response to von Willebrand factor and thrombin. Eur J Biochem. 1985 Nov 15;153(1):1–11. doi: 10.1111/j.1432-1033.1985.tb09259.x. [DOI] [PubMed] [Google Scholar]
  42. Zafar R. S., Walz D. A. Platelet membrane glycoprotein V: characterization of the thrombin-sensitive glycoprotein from human platelets. Thromb Res. 1989 Jan 1;53(1):31–44. doi: 10.1016/0049-3848(89)90113-8. [DOI] [PubMed] [Google Scholar]
  43. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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