Skip to main content
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1984 Feb;73(2):291–297. doi: 10.1172/JCI111213

An altered platelet granule glycoprotein in patients with essential thrombocythemia.

W J Booth, M C Berndt, P A Castaldi
PMCID: PMC425018  PMID: 6365970

Abstract

The protein profiles of washed platelets from nine patients with essential thrombocythemia were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In four patients, an additional protein band (reduced Mr of 170,000) was clearly identified in both unstimulated platelet preparations and thrombin-released supernatant fractions. This band was also evident, though to a lesser extent, in three more patients, but it could not be located in the two remaining patients nor in any of ten controls. Subsequent characterization of the 170,000 reduced protein in one patient indicated that (a) it was glycosylated, as judged by periodic acid-Schiff staining, and (b) that native protein was a disulfide-linked multimer (possibly trimeric), which (c) partially bound to the activated platelet plasma membrane in the presence of calcium, and (d) was immune precipitated by anti-glycoprotein G antisera. The combined evidence is consistent with the 170,000 reduced protein being a modified form of the normal subunit of the platelet alpha-granule constituent, glycoprotein G (also termed thrombospondin and thrombin-sensitive protein).

Full text

PDF

Images in this article

Selected References

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

  1. Adams T., Schutz L., Goldberg L. Platelet function abnormalities in the myeloproliferative disorders. Scand J Haematol. 1974;13(3):215–224. doi: 10.1111/j.1600-0609.1974.tb00262.x. [DOI] [PubMed] [Google Scholar]
  2. Baenziger N. L., Brodie G. N., Majerus P. W. Isolation and properties of a thrombin-sensitive protein of human platelets. J Biol Chem. 1972 May 10;247(9):2723–2731. [PubMed] [Google Scholar]
  3. Berndt M. C., Castaldi P. A., Gordon S., Halley H., McPherson V. J. Morphological and biochemical confirmation of gray platelet syndrome in two siblings. Aust N Z J Med. 1983 Aug;13(4):387–390. doi: 10.1111/j.1445-5994.1983.tb04488.x. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Bolin R. B., Okumura T., Jamieson G. A. Changes in distribution of platelet membrane glycoproteins in patients with myeloproliferative disorders. Am J Hematol. 1977;3:63–71. doi: 10.1002/ajh.2830030108. [DOI] [PubMed] [Google Scholar]
  6. Boneu B., Nouvel C., Sie P., Caranobe C., Combes D., Laurent G., Pris J., Bierme R. Platelets in myeloproliferative disorders. I. A comparative evaluation with certain platelet function tests. Scand J Haematol. 1980 Sep;25(3):214–220. [PubMed] [Google Scholar]
  7. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  8. Caranobe C., Sie P., Nouvel C., Laurent G., Pris J., Boneu B. Platelets in myeloproliferative disorders. II. Serotonin uptake and storage: correlations with mepacrine labelled dense bodies and with platelet density. Scand J Haematol. 1980 Oct;25(4):289–295. [PubMed] [Google Scholar]
  9. Castaldi P. A., Berndt M. C., Booth W., Gregory C., Bull H., Greaves M. Evidence for a platelet membrane defect in the myeloproliferative syndromes. Thromb Res. 1982 Sep 1;27(5):601–609. doi: 10.1016/0049-3848(82)90307-3. [DOI] [PubMed] [Google Scholar]
  10. Cooper B., Schafer A. I., Puchalsky D., Handin R. I. Platelet resistance to prostaglandin D2 in patients with myeloproliferative disorders. Blood. 1978 Sep;52(3):618–626. [PubMed] [Google Scholar]
  11. Eche N., Sie P., Caranobe C., Nouvel C., Pris J., Boneu B. Platelets in myeloproliferative disorders. III: Glycoprotein profile in relation to platelet function and platelet density. Scand J Haematol. 1981 Feb;26(2):123–129. doi: 10.1111/j.1600-0609.1981.tb01634.x. [DOI] [PubMed] [Google Scholar]
  12. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  13. Gartner T. K., Gerrard J. M., White J. G., Williams D. C. Fibrinogen is the receptor for the endogenous lectin of human platelets. Nature. 1981 Feb 19;289(5799):688–690. doi: 10.1038/289688a0. [DOI] [PubMed] [Google Scholar]
  14. Gartner T. K., Gerrard J. M., White J. G., Williams D. C. The endogenous lectin of human platelets is an alpha-granule component. Blood. 1981 Jul;58(1):153–157. [PubMed] [Google Scholar]
  15. Gartner T. K., Williams D. C., Minion F. C., Phillips D. R. Thrombin-induced platelet aggregation is mediated by a platelet plasma membrane-bound lectin. Science. 1978 Jun 16;200(4347):1281–1283. doi: 10.1126/science.663608. [DOI] [PubMed] [Google Scholar]
  16. George J. N. Studies on platelet plasma membranes. IV. Quantitative analysis of platelet membrane glycoproteins by (125I)-diazotized diiodosulfanilic acid labeling and SDS-polyacrylamide gel electrophoresis. J Lab Clin Med. 1978 Sep;92(3):430–446. [PubMed] [Google Scholar]
  17. Gerrard J. M., Phillips D. R., Rao G. H., Plow E. F., Walz D. A., Ross R., Harker L. A., White J. G. Biochemical studies of two patients with the gray platelet syndrome. Selective deficiency of platelet alpha granules. J Clin Invest. 1980 Jul;66(1):102–109. doi: 10.1172/JCI109823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ginsburg A. D. Platelet function in patients with high platelet counts. Ann Intern Med. 1975 Apr;82(4):506–511. doi: 10.7326/0003-4819-82-4-506. [DOI] [PubMed] [Google Scholar]
  19. HARDISTY R. M., WOLFF H. H. Haemorrhagic thrombocythaemia: a clinical and laboratory study. Br J Haematol. 1955 Oct;1(4):390–405. doi: 10.1111/j.1365-2141.1955.tb05528.x. [DOI] [PubMed] [Google Scholar]
  20. Hawkes R., Niday E., Gordon J. A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem. 1982 Jan 1;119(1):142–147. doi: 10.1016/0003-2697(82)90677-7. [DOI] [PubMed] [Google Scholar]
  21. Jaffe E. A., Leung L. L., Nachman R. L., Levin R. I., Mosher D. F. Thrombospondin is the endogenous lectin of human platelets. Nature. 1982 Jan 21;295(5846):246–248. doi: 10.1038/295246a0. [DOI] [PubMed] [Google Scholar]
  22. Jaffe E. A., Ruggiero J. T., Leung L. K., Doyle M. J., McKeown-Longo P. J., Mosher D. F. Cultured human fibroblasts synthesize and secrete thrombospondin and incorporate it into extracellular matrix. Proc Natl Acad Sci U S A. 1983 Feb;80(4):998–1002. doi: 10.1073/pnas.80.4.998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Keenan J. P., Wharton J., Shepherd A. J., Bellingham A. J. Defective platelet lipid peroxidation in myeloproliferative disorders: a possible defect of prostaglandin synthesis. Br J Haematol. 1977 Feb;35(2):275–283. doi: 10.1111/j.1365-2141.1977.tb00584.x. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Lawler J. W., Chao F. C., Fang P. H. Observation of a high molecular weight platelet protein released by thrombin. Thromb Haemost. 1977 Apr 30;37(2):355–357. [PubMed] [Google Scholar]
  26. Lawler J. W., Slayter H. S., Coligan J. E. Isolation and characterization of a high molecular weight glycoprotein from human blood platelets. J Biol Chem. 1978 Dec 10;253(23):8609–8616. [PubMed] [Google Scholar]
  27. Lawler J., Chao F. C., Cohen C. M. Evidence for calcium-sensitive structure in platelet thrombospondin. Isolation and partial characterization of thrombospondin in the presence of calcium. J Biol Chem. 1982 Oct 25;257(20):12257–12265. [PubMed] [Google Scholar]
  28. Leung L. L., Nachman R. L. Complex formation of platelet thrombospondin with fibrinogen. J Clin Invest. 1982 Sep;70(3):542–549. doi: 10.1172/JCI110646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mosher D. F., Doyle M. J., Jaffe E. A. Synthesis and secretion of thrombospondin by cultured human endothelial cells. J Cell Biol. 1982 May;93(2):343–348. doi: 10.1083/jcb.93.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nurden A. T., Kunicki T. J., Dupuis D., Soria C., Caen J. P. Specific protein and glycoprotein deficiencies in platelets isolated from two patients with the gray platelet syndrome. Blood. 1982 Apr;59(4):709–718. [PubMed] [Google Scholar]
  31. Pareti F. I., Gugliotta L., Mannucci L., Guarini A., Mannucci P. M. Biochemical and metabolic aspects of platelet dysfunction in chronic myeloproliferative disorders. Thromb Haemost. 1982 Apr 30;47(2):84–89. [PubMed] [Google Scholar]
  32. 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]
  33. Phillips D. R., Jennings L. K., Prasanna H. R. Ca2+-mediated association of glycoprotein G (thrombinsensitive protein, thrombospondin) with human platelets. J Biol Chem. 1980 Dec 25;255(24):11629–11632. [PubMed] [Google Scholar]
  34. Saglio S. D., Slayter H. S. Use of a radioimmunoassay to quantify thrombospondin. Blood. 1982 Jan;59(1):162–166. [PubMed] [Google Scholar]
  35. Schafer A. I. Deficiency of platelet lipoxygenase activity in myeloproliferative disorders. N Engl J Med. 1982 Feb 18;306(7):381–386. doi: 10.1056/NEJM198202183060701. [DOI] [PubMed] [Google Scholar]
  36. Spaet T. H., Gaynor E., Goldstein M. L. Defective platelets in essential thrombocythemia. Arch Intern Med. 1969 Aug;124(2):135–141. [PubMed] [Google Scholar]
  37. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem. 1969 Jul;30(1):148–152. doi: 10.1016/0003-2697(69)90383-2. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES