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Thromb Haemost 1994; 71(04): 493-498
DOI: 10.1055/s-0038-1642466
Review Article
Schattauer GmbH Stuttgart

Demonstration of a Functionally Active tPA-Like Plasminogen Activator in Human Platelets

D L Wang
The Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, ROC
,
Y T Pan
The Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, ROC
,
J J Wang
The Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, ROC
,
C H Cheng
The Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, ROC
,
C Y Liu
The Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, ROC
› Author Affiliations
Further Information

Publication History

Received: 03 March 1993

Accepted after resubmission: 23 December 1993

Publication Date:
06 July 2018 (online)

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Summary

The mechanism of platelet-enhanced fibrinolysis is unclear. We therefore investigated the fibrinolytic activity of human platelets and demonstrated that they contain a tissue plasminogen activator (tPA)- like plasminogen activator, abbreviated as tPA-like-PA. This activator was detected by ELISA in platelet incubation medium and in platelet Triton extracts. Plasminogen activation assays showed that this tPA- like-PA could induce plasminogen activation to form plasmin. Western blots of Triton extracts incubated with anti-tPA antibody demonstrated a major 64-kD protein band, compared to a 70-kD band for standard single chain tPA, plus a minor 118-kD band corresponding to a complex of tPA-like-PA and plasminogen activator inhibitor (PAI-1). Western blots of Triton extracts incubated with anti-PAI-1 antibody produced an approximately similar high-molecular-weight (118 kD) protein band. Fibrin zymographic analysis of affinity-purified tPA-like- PA demonstrated a major and a minor fibrin lysis zone, which approximately corresponded to the tPA-like-PA and its complex with PAI-1 observed by Western blots. Immunogold labelling and electron microscopy demonstrated that platelet activator, either as the free form or co-localized with PAI-1, was present in granules and in channels of the open canalicular system. We conclude that platelets contain a functionally active tPA-like-PA, whose low fibrinolytic activity might be due to its readily forming a complex with PAI-1. This functionally active tPA-like-PA might contribute to the enhanced fibrinolytic activity of platelets observed in platelet-rich thrombi.

 

  • References

  • 1 Erikson LA, Ginsberg MH, Loskutoff DJ. Detection and partial characterization of an inhibitor of plasminogen activator in human platelets. J Clin Invest 1984; 74: 1465-1468
    CrossrefPubMedSearch in Google Scholar
  • 2 Plow EF, Collen D. The presence and release of alpha-2 antiplasmin from human platelets. Blood 1981; 58: 1069-1071
    CrossrefPubMedSearch in Google Scholar
  • 3 Schwartz ML, Pizzo V, Hill RL, McKee PA. Human factor XIII from plasma and platelets: molecular weight, subunit structures, proteolytic activation and cross-linking of fibrinogen and fibrin. J Biol Chem 1973; 248: 1395-407
    PubMedSearch in Google Scholar
  • 4 Reid WO, Silver MJ. Activation of the blood fibrinolytic enzyme system by platelets. Am J Physiol 1964; 206: 1255-1260
    PubMedSearch in Google Scholar
  • 5 Lockhart MS, Comp PC, Taylor Jr FB. Role of platelets in lysis of dilute plasma clots. J Lab Clin Med 1979; 94: 285-290
    PubMedSearch in Google Scholar
  • 6 Carrol RC, Radcliffe RD, Taylor Jr FB. Plasminogen, plasminogen activator and platelets in the regulation of clot lysis: reconstitute and patient, studies. J Lab Clin Med 1982; 100: 986-990
    PubMedSearch in Google Scholar
  • 7 Deguchi K, Murashima S, Shirakawa S, Soria C, Soria J, Dunn F, Tobelem G. The potentiation effect of platelets on plasminogen activation by tissue plasminogen activator. Thromb Res 1985; 49: 853-861
    PubMedSearch in Google Scholar
  • 8 Miles LA, Plow EF. Binding and activation of plasminogen on the platelet surface. J Biol Chem 1985; 206 (07) 4303-4311
    PubMedSearch in Google Scholar
  • 9 Vaughan DE, Mendelsohn ME, Declerck PJ, Van HoutteE, Collen D, Los-calzo J. Characterization of the binding of human tissue-type plasminogen activator to platelets. J Biol Chem 1989; 264 (27) 15869-15874
    CrossrefPubMedSearch in Google Scholar
  • 10 Park S, Harker LA, Marzec UM, Levin EG. Demonstration of single chain urokinase-type plasminogen activator on human platelet membrane. Blood 1989; 73 (06) 1421-1425
    PubMedSearch in Google Scholar
  • 11 Jeanneau C, Sultan Y. Tissue plasminogen activator in human megakaryocytes and platelets: immunocytochemical localization, immunoblotting and zymographic analysis. Thromb Haemostas 1988; 59: 529-534
    PubMedSearch in Google Scholar
  • 12 Wang DL, Chang YN, Hsu HTC, Usami S, Chien S. Prostaglandin Ej and dibutyryl cyclic AMP enhance platelet resistance to deformation. Thromb Res 1992; 65: 757-768
    CrossrefPubMedSearch in Google Scholar
  • 13 Huang TF, Sheu JR, Teng CM, Chen SW, Liu CS. Triflavin, an antiplatelet Arg-Gly-Asp-containing peptide, is a specific antagonist of platelet membrane glycoprotein Ilb-IIIa complex. J Biochem 1991; 109: 328-334
    PubMedSearch in Google Scholar
  • 14 Wallen P, Pohl G, Bergsdorf N, Randy M, Ny T, Jornvall H. Purification and characterization of a melanoma cell plasminogen activator. Eur J Biochem 1983; 132: 681-686
    CrossrefPubMedSearch in Google Scholar
  • 15 Morris JP, Blatt S, Powell JR, Strickland DK, Castellino FJ. Role of lysine binding regions in the kinetic properties of human plasmin. Biochem 1981; 20: 4811-4816
    CrossrefPubMedSearch in Google Scholar
  • 16 Liu CY, Wallen P, Handley DA, Smith J. In: Fibrinogen and its Derivatives: Biochemistry Physiology and Pathophysiology. Muller BG. et al. eds. Excerpta Medica; New York: 1986: 319-322
  • 17 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1979; 227: 680-685
    PubMedSearch in Google Scholar
  • 18 Wang DL, Annamalai AE, Ghosh S, Gewirtz AM, Colman RW. Human platelet factor V is crosslinked to actin by factor XHIa during platelet activation by thrombin. Thromb Res 1990; 57: 39-57
    CrossrefPubMedSearch in Google Scholar
  • 19 Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gel to nitrocellulose sheets: procedures and some applications. Proc Natl Acad Sci (USA) 1979; 76: 4350-4356
    CrossrefPubMedSearch in Google Scholar
  • 20 Levin EG, Loskutoff DJ. Cultured bovine endothelial cells produce both urokinase and tissue-type plasminogen activator. J Cell Biol 1982; 94: 631-636
    CrossrefPubMedSearch in Google Scholar
  • 21 Wang JJ, Wang DL. Immuno-electron microscopic studies of endothelium in cardiovascular system. J Formosan Med Assoc 1992; 91: S262-9
    PubMedSearch in Google Scholar
  • 22 Harrison P, Wilbourn B, Debili N, Vainchenker W, Breton Gorius J, Lawrie AS, Masse JM, Savidge GF, Cramer EM. Uptake of plasma fibrinogen into alpha granules of human megakaryocytes and platelets. J Clin Invest 1989; 84: 1320-1324
    CrossrefPubMedSearch in Google Scholar
  • 23 White JG, Escolar G. The blood platelet open canalicular system: a two- way street. Eur J Cell Biol 1991; 56: 233-242
    PubMedSearch in Google Scholar
  • 24 Behnke O. Coated pits and vesicles transfer plasma components to platelet granules. Thromb Haemostas 1989; 62: 718-722
    PubMedSearch in Google Scholar
  • 25 Morgenstern E, Ruf A, Patscheke H. Transport of antiglycoprotein IIb/IIIa- antibody into the alpha-granules of unstimulated human blood platelets. Thromb Haemostas 1992; 67: 121-125
    PubMedSearch in Google Scholar
  • 26 Both NA, Simpson AJ, Croll A, Bennet B, MacGregor IR. Plasminogen activator inhibitor (PAI-1) in plasma and platelets. Br J Haematol 1988; 70: 327-333
    CrossrefPubMedSearch in Google Scholar
  • 27 Rijken DC, Juhan-Vague I, De Cock F, Collen D. Measurement of human tissue-type plasminogen activator by a two-site immunoradiometric assay. J Lab Clin Med 1983; 101: 274
    PubMedSearch in Google Scholar
  • 28 Underhill DM, Owensby DA, Morton PA, Schwartz AL. Endocytosis and lysosomal delivery of tissue plasminogen activator-inhibitor 1 complexes in Hep G2 cells. Blood 1992; 80 (11) 2746-2754
    PubMedSearch in Google Scholar