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. 1994 Jun;93(6):2417–2424. doi: 10.1172/JCI117249

Complementary DNA cloning of the alternatively expressed endothelial cell glycoprotein Ib beta (GPIb beta) and localization of the GPIb beta gene to chromosome 22.

M D Kelly 1, D W Essex 1, S S Shapiro 1, F J Meloni 1, T Druck 1, K Huebner 1, B A Konkle 1
PMCID: PMC294449  PMID: 8200976

Abstract

Glycoprotein Ib beta (GPIb beta) exists in platelets disulfide-linked to glycoprotein Ib alpha (GPIb alpha), a major receptor for von Willebrand factor. Both GPIb alpha and GPIb beta are expressed in endothelial cells (EC). While the GPIb alpha mRNA and protein appear similar in platelets and EC, EC GPIb beta mRNA is larger than platelet GPIb beta and encodes a larger protein. We have cloned and sequenced EC GPIb beta cDNA and report a 2793-nucleotide sequence which contains a 411-amino acid open reading frame. The EC sequence contains all of the platelet cDNA sequence and all but three amino acids of the primary translation product. Like the genes encoding GPIb alpha, GPIX, and GPV, the GPIb beta gene appears simple in structure. Using human hamster hybrids, we have localized the GPIb beta gene to chromosome 22pter-->22q11.2. When we examined poly (A)+ RNA from several human tissues for GPIb beta mRNA expression, we found that GPIb beta mRNA was expressed in a variety of tissues but was most abundant in heart and brain, while GPIb alpha and GPIX mRNA expression was found only in lung and placenta at very low levels. The broad distribution of GPIb beta mRNA suggests that it may be playing a role different than or additional to its function in platelets.

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Selected References

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

  1. Adams M. D., Dubnick M., Kerlavage A. R., Moreno R., Kelley J. M., Utterback T. R., Nagle J. W., Fields C., Venter J. C. Sequence identification of 2,375 human brain genes. Nature. 1992 Feb 13;355(6361):632–634. doi: 10.1038/355632a0. [DOI] [PubMed] [Google Scholar]
  2. Andrews R. K., Fox J. E. Interaction of purified actin-binding protein with the platelet membrane glycoprotein Ib-IX complex. J Biol Chem. 1991 Apr 15;266(11):7144–7147. [PubMed] [Google Scholar]
  3. Asch A. S., Adelman B., Fujimoto M., Nachman R. L. Identification and isolation of a platelet GPIb-like protein in human umbilical vein endothelial cells and bovine aortic smooth muscle cells. J Clin Invest. 1988 May;81(5):1600–1607. doi: 10.1172/JCI113494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bauer S. R., Huebner K., Budarf M., Finan J., Erikson J., Emanuel B. S., Nowell P. C., Croce C. M., Melchers F. The human Vpre B gene is located on chromosome 22 near a cluster of V lambda gene segments. Immunogenetics. 1988;28(5):328–333. doi: 10.1007/BF00364231. [DOI] [PubMed] [Google Scholar]
  5. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  6. Chou P. Y., Fasman G. D. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. doi: 10.1002/9780470122921.ch2. [DOI] [PubMed] [Google Scholar]
  7. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  8. Fugman D. A., Witte D. P., Jones C. L., Aronow B. J., Lieberman M. A. In vitro establishment and characterization of a human megakaryoblastic cell line. Blood. 1990 Mar 15;75(6):1252–1261. [PubMed] [Google Scholar]
  9. Garnier J., Osguthorpe D. J., Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. doi: 10.1016/0022-2836(78)90297-8. [DOI] [PubMed] [Google Scholar]
  10. Ginsburg D., Handin R. I., Bonthron D. T., Donlon T. A., Bruns G. A., Latt S. A., Orkin S. H. Human von Willebrand factor (vWF): isolation of complementary DNA (cDNA) clones and chromosomal localization. Science. 1985 Jun 21;228(4706):1401–1406. doi: 10.1126/science.3874428. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Hickey M. J., Hagen F. S., Yagi M., Roth G. J. Human platelet glycoprotein V: characterization of the polypeptide and the related Ib-V-IX receptor system of adhesive, leucine-rich glycoproteins. Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8327–8331. doi: 10.1073/pnas.90.18.8327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Huebner K., Druck T., Croce C. M., Thiesen H. J. Twenty-seven nonoverlapping zinc finger cDNAs from human T cells map to nine different chromosomes with apparent clustering. Am J Hum Genet. 1991 Apr;48(4):726–740. [PMC free article] [PubMed] [Google Scholar]
  15. Khan A. S., Wilcox A. S., Polymeropoulos M. H., Hopkins J. A., Stevens T. J., Robinson M., Orpana A. K., Sikela J. M. Single pass sequencing and physical and genetic mapping of human brain cDNAs. Nat Genet. 1992 Nov;2(3):180–185. doi: 10.1038/ng1192-180. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Konkle B. A., Ginsburg D. The addition of endothelial cell growth factor and heparin to human umbilical vein endothelial cell cultures decreases plasminogen activator inhibitor-1 expression. J Clin Invest. 1988 Aug;82(2):579–585. doi: 10.1172/JCI113635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987 Oct 26;15(20):8125–8148. doi: 10.1093/nar/15.20.8125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kozak M. An analysis of vertebrate mRNA sequences: intimations of translational control. J Cell Biol. 1991 Nov;115(4):887–903. doi: 10.1083/jcb.115.4.887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  21. Kozak M. Regulation of translation in eukaryotic systems. Annu Rev Cell Biol. 1992;8:197–225. doi: 10.1146/annurev.cb.08.110192.001213. [DOI] [PubMed] [Google Scholar]
  22. Lanza F., Morales M., de La Salle C., Cazenave J. P., Clemetson K. J., Shimomura T., Phillips D. R. Cloning and characterization of the gene encoding the human platelet glycoprotein V. A member of the leucine-rich glycoprotein family cleaved during thrombin-induced platelet activation. J Biol Chem. 1993 Oct 5;268(28):20801–20807. [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. 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]
  26. 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]
  27. McKeown M. Alternative mRNA splicing. Annu Rev Cell Biol. 1992;8:133–155. doi: 10.1146/annurev.cb.08.110192.001025. [DOI] [PubMed] [Google Scholar]
  28. Miller J. L., Lyle V. A., Cunningham D. Mutation of leucine-57 to phenylalanine in a platelet glycoprotein Ib alpha leucine tandem repeat occurring in patients with an autosomal dominant variant of Bernard-Soulier disease. Blood. 1992 Jan 15;79(2):439–446. [PubMed] [Google Scholar]
  29. 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]
  30. Rajagopalan V., Essex D. W., Shapiro S. S., Konkle B. A. Tumor necrosis factor-alpha modulation of glycoprotein Ib alpha expression in human endothelial and erythroleukemia cells. Blood. 1992 Jul 1;80(1):153–161. [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. Ruggeri Z. M. The platelet glycoprotein Ib-IX complex. Prog Hemost Thromb. 1991;10:35–68. [PubMed] [Google Scholar]
  34. 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]
  35. 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]
  36. 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]
  37. Ware J., Russell S. R., Vicente V., Scharf R. E., Tomer A., McMillan R., Ruggeri Z. M. Nonsense mutation in the glycoprotein Ib alpha coding sequence associated with Bernard-Soulier syndrome. Proc Natl Acad Sci U S A. 1990 Mar;87(5):2026–2030. doi: 10.1073/pnas.87.5.2026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Wright S. D., Michaelides K., Johnson D. J., West N. C., Tuddenham E. G. Double heterozygosity for mutations in the platelet glycoprotein IX gene in three siblings with Bernard-Soulier syndrome. Blood. 1993 May 1;81(9):2339–2347. [PubMed] [Google Scholar]
  40. Wyler B., Bienz D., Clemetson K. J., Luscher E. F. Glycoprotein Ib beta is the only phosphorylated major membrane glycoprotein in human platelets. Biochem J. 1986 Mar 1;234(2):373–379. doi: 10.1042/bj2340373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985 Jul 5;184(1):99–105. doi: 10.1016/0022-2836(85)90046-4. [DOI] [PubMed] [Google Scholar]
  42. von Heijne G. The signal peptide. J Membr Biol. 1990 May;115(3):195–201. doi: 10.1007/BF01868635. [DOI] [PubMed] [Google Scholar]

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