Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1986 Mar;77(3):750–756. doi: 10.1172/JCI112370

Exposure of fibrinogen receptors in human platelets by surface proteolysis with elastase.

E Kornecki, Y H Ehrlich, D D De Mars, R H Lenox
PMCID: PMC423459  PMID: 3005363

Abstract

Human platelets that were preincubated with porcine elastase aggregated spontaneously upon the addition of fibrinogen. Maximal aggregation to fibrinogen was observed with platelets pretreated with an elastase concentration of 111 micrograms/ml, and half-maximal aggregation occurred after treatment with 11 micrograms/ml elastase. Binding of radiolabeled fibrinogen to elastase-treated platelets was specific, saturable, and showed a single class of 48,400 +/- 9,697 fibrinogen-binding sites per platelet with a dissociation constant of 6.30 +/- 1.48 X 10(-7) M. ATP, apyrase, and the stimulators of platelet adenylate cyclase forskolin, prostaglandin E1, prostacyclin, and N6, 2'-O-dibutyryl cyclic AMP did not inhibit the fibrinogen-induced aggregation of elastase-treated platelets. EDTA completely blocked the initiation of aggregation and reversed the fibrinogen-induced aggregation of elastase-treated platelets. Monoclonal and polyclonal antibodies directed against glycoproteins (GP) IIb and IIIa completely blocked the fibrinogen-induced aggregation of elastase-treated platelets. Immunoprecipitates with these antibodies obtained from detergent extracts of surface-radiolabeled, intact, and elastase-treated platelets contained the glycoproteins IIb and IIIa. We conclude that surface proteolysis by low concentrations of elastase can expose fibrinogen-binding sites associated with GPIIb and GPIIIa on the platelet surface, resulting in spontaneous aggregation upon the addition of fibrinogen. These findings may be relevant to hemostatic changes observed in patients with increased levels of circulating elastase.

Full text

PDF
750

Images in this article

753Image
on p.753
754Image
on p.754

Selected References

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

  1. BALO J., BANGA I. Change in the elastase content of the pancreas in relation to arteriosclerosis. Acta Physiol Acad Sci Hung. 1953;4(1-2):187–194. [PubMed] [Google Scholar]
  2. BRINK N. G., LEWIS U. J., WILLIAMS D. E. Pancreatic elastase: purification, properties, and function. J Biol Chem. 1956 Oct;222(2):705–720. [PubMed] [Google Scholar]
  3. Bennett J. S., Vilaire G., Burch J. W. A role for prostaglandins and thromboxanes in the exposure of platelet fibrinogen receptors. J Clin Invest. 1981 Oct;68(4):981–987. doi: 10.1172/JCI110352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bennett J. S., Vilaire G., Cines D. B. Identification of the fibrinogen receptor on human platelets by photoaffinity labeling. J Biol Chem. 1982 Jul 25;257(14):8049–8054. [PubMed] [Google Scholar]
  5. 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]
  6. Brower M. S., Levin R. I., Garry K. Human neutrophil elastase modulates platelet function by limited proteolysis of membrane glycoproteins. J Clin Invest. 1985 Feb;75(2):657–666. doi: 10.1172/JCI111744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Egbring R., Schmidt W., Fuchs G., Havemann K. Demonstration of granulocytic proteases in plasma of patients with acute leukemia and septicemia with coagulation defects. Blood. 1977 Feb;49(2):219–231. [PubMed] [Google Scholar]
  8. Favis G. R., Colman R. W. The action of halofenate on platelet shape change and prostaglandin synthesis. J Lab Clin Med. 1978 Jul;92(1):45–52. [PubMed] [Google Scholar]
  9. Greenberg J. P., Packham M. A., Guccione M. A., Harfenist E. J., Orr J. L., Kinlough-Rathbone R. L., Perry D. W., Mustard J. F. The effect of pretreatment of human or rabbit platelets with chymotrypsin on their responses to human fibrinogen and aggregating agents. Blood. 1979 Oct;54(4):753–765. [PubMed] [Google Scholar]
  10. Hagen I., Nurden A., Bjerrum O. J., Solum N. O., Caen J. Immunochemical evidence for protein abnormalities in platelets from patients with Glanzmann's thrombasthenia and Bernard-Soulier syndrome. J Clin Invest. 1980 Mar;65(3):722–731. doi: 10.1172/JCI109719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hantgan R. R. A study of the kinetics of ADP-triggered platelet shape change. Blood. 1984 Oct;64(4):896–906. [PubMed] [Google Scholar]
  12. Harpel P. C., Mosesson M. W. Degradation of human fibrinogen by plasms alpha2-macroglobulin-enzyme complexes. J Clin Invest. 1973 Sep;52(9):2175–2184. doi: 10.1172/JCI107402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hawiger J., Parkinson S., Timmons S. Prostacyclin inhibits mobilisation of fibrinogen-binding sites on human ADP- and thrombin-treated platelets. Nature. 1980 Jan 10;283(5743):195–197. doi: 10.1038/283195a0. [DOI] [PubMed] [Google Scholar]
  14. Kopeć M., Bykowska K., Lopaciuk S., Jelenska M., Kaczanowska J., Sopata I., Wojtecka E. Effects of neutral proteases from human leukocytes on structure and biological properties of human factor VIII. Thromb Haemost. 1980 Jul 15;43(3):211–217. [PubMed] [Google Scholar]
  15. Kornecki E., Lee H., Merlin F., Hershock D., Tuszynski G. P., Niewiarowski S. Comparison of platelet fibrinogen receptors on intact and proteolytically-treated platelets by use of an anti-glycoprotein IIIa monoclonal antibody (MA 123). Thromb Res. 1984 Apr 1;34(1):35–49. doi: 10.1016/0049-3848(84)90104-x. [DOI] [PubMed] [Google Scholar]
  16. Kornecki E., Niewiarowski S., Morinelli T. A., Kloczewiak M. Effects of chymotrypsin and adenosine diphosphate on the exposure of fibrinogen receptors on normal human and Glanzmann's thrombasthenic platelets. J Biol Chem. 1981 Jun 10;256(11):5696–5701. [PubMed] [Google Scholar]
  17. Kornecki E., Tuszynski G. P., Niewiarowski S. Inhibition of fibrinogen receptor-mediated platelet aggregation by heterologous anti-human platelet membrane antibody. Significance of an Mr = 66,000 protein derived from glycoprotein IIIa. J Biol Chem. 1983 Aug 10;258(15):9349–9356. [PubMed] [Google Scholar]
  18. Lenox R. H., Ellis J., Van Riper D., Ehrlich Y. H. Alpha 2-adrenergic receptor-mediated regulation of adenylate cyclase in the intact human platelet. Evidence for a receptor reserve. Mol Pharmacol. 1985 Jan;27(1):1–9. [PubMed] [Google Scholar]
  19. 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]
  20. McEver R. P., Bennett E. M., Martin M. N. Identification of two structurally and functionally distinct sites on human platelet membrane glycoprotein IIb-IIIa using monoclonal antibodies. J Biol Chem. 1983 Apr 25;258(8):5269–5275. [PubMed] [Google Scholar]
  21. Milton J. G., Yung W., Glushak C., Frojmovic M. M. Kinetics of ADP-induced human platelet shape change: apparent positive cooperativity. Can J Physiol Pharmacol. 1980 Jan;58(1):45–52. doi: 10.1139/y80-009. [DOI] [PubMed] [Google Scholar]
  22. Morinelli T. A., Niewiarowski S., Kornecki E., Figures W. R., Wachtfogel Y., Colman R. W. Platelet aggregation and exposure of fibrinogen receptors by prostaglandin endoperoxide analogues. Blood. 1983 Jan;61(1):41–49. [PubMed] [Google Scholar]
  23. Mustard J. F., Perry D. W., Ardlie N. G., Packham M. A. Preparation of suspensions of washed platelets from humans. Br J Haematol. 1972 Feb;22(2):193–204. doi: 10.1111/j.1365-2141.1972.tb08800.x. [DOI] [PubMed] [Google Scholar]
  24. Nachman R. L., Leung L. L. Complex formation of platelet membrane glycoproteins IIb and IIIa with fibrinogen. J Clin Invest. 1982 Feb;69(2):263–269. doi: 10.1172/JCI110448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Niewiarowski S., Budzynski A. Z., Morinelli T. A., Brudzynski T. M., Stewart G. J. Exposure of fibrinogen receptor on human platelets by proteolytic enzymes. J Biol Chem. 1981 Jan 25;256(2):917–925. [PubMed] [Google Scholar]
  26. Ordinas A., Maragall S., Castillo R., Nurden A. T. A glycoprotein I defect in the platelets of three patients with severe cirrhosis of the liver. Thromb Res. 1978 Aug;13(2):297–302. doi: 10.1016/0049-3848(78)90018-x. [DOI] [PubMed] [Google Scholar]
  27. Peerschke E. I., Zucker M. B., Grant R. A., Egan J. J., Johnson M. M. Correlation between fibrinogen binding to human platelets and platelet aggregability. Blood. 1980 May;55(5):841–847. [PubMed] [Google Scholar]
  28. Peerschke E. I., Zucker M. B. Relationship of ADP-induced fibrinogen binding to platelet shape change and aggregation elucidated by use of colchicine and cytochalasin B. Thromb Haemost. 1980 Feb 29;43(1):58–60. [PubMed] [Google Scholar]
  29. 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]
  30. Plow E. F. Leukocyte elastase release during blood coagulation. A potential mechanism for activation of the alternative fibrinolytic pathway. J Clin Invest. 1982 Mar;69(3):564–572. doi: 10.1172/JCI110482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Plow E. F., Marguerie G. A. Induction of the fibrinogen receptor on human platelets by epinephrine and the combination of epinephrine and ADP. J Biol Chem. 1980 Nov 25;255(22):10971–10977. [PubMed] [Google Scholar]
  32. Plow E. F., Marguerie G. A. Participation of ADP in the binding of fibrinogen to thrombin-stimulated platelets. Blood. 1980 Sep;56(3):553–555. [PubMed] [Google Scholar]
  33. Prinz R. A., Fareed J., Rock A., Squillaci G., Wallenga J. Platelet activation by human pancreatic fluid. J Surg Res. 1984 Oct;37(4):314–319. doi: 10.1016/0022-4804(84)90195-1. [DOI] [PubMed] [Google Scholar]
  34. Salomon Y. Adenylate cyclase assay. Adv Cyclic Nucleotide Res. 1979;10:35–55. [PubMed] [Google Scholar]
  35. Sterrenberg L., Gravesen M., Haverkate F., Nieuwenhuizen W. Granulocyte enzyme mediated degradation of human fibrinogen in plasma in vitro. Thromb Res. 1983 Sep 1;31(5):719–728. doi: 10.1016/0049-3848(83)90102-0. [DOI] [PubMed] [Google Scholar]
  36. Switzer M. E., Gordon H. J., McKee P. A. Proteolytic activity of alpha 2-macroglobulin-enzyme complexes toward human factor VIII/von Willebrand factor. Biochemistry. 1983 Mar 15;22(6):1437–1444. doi: 10.1021/bi00275a018. [DOI] [PubMed] [Google Scholar]
  37. Wheeler M. E., Cox A. C., Carroll R. C. Retention of the glycoprotein IIb-IIIa complex in the isolated platelet cytoskeleton. Effects of separable assembly of platelet pseudopodal and contractile cytoskeletons. J Clin Invest. 1984 Sep;74(3):1080–1089. doi: 10.1172/JCI111475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Whittemore S. R., Graber S. G., Lenox R. H., Hendley E. D., Ehrlich Y. H. Activation of adenylyl cyclase by preincubation of rat cerebral-cortical membranes under phosphorylating conditions: role of ATP, GTP, and divalent cations. J Neurochem. 1984 Jun;42(6):1685–1696. doi: 10.1111/j.1471-4159.1984.tb12760.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES