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DOI: 10.1055/s-0038-1653843
Exposure of Human Platelets to Plasmin Results in the Expression of Irreversibly Active Fibrinogen Receptors
Publication History
Received 31 August 1994
Accepted after resubmission 10 January 1995
Publication Date:
09 July 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199504/lookinside/thumbnails/10-1055-s-0038-1653843-1.jpg)
Summary
Although plasmin can trigger strong platelet responses such as shape change and exocytosis of internal granules, limited platelet aggregation is induced by this proteinase, owing to its capacity to rapidly proteolyse secreted adhesive proteins. In this context, we have investigated the state of activation of the fibrinogen receptor, the integrin αIIbβ3, on platelets exposed to plasmin. Following incubation with plasmin at 37 °C, washing, and resuspension, platelets exhibit a moderate, low-velocity aggregation when stirred in the presence of fibrinogen. Optimum aggregability is observed when platelets have been exposed to plasmin activity of ≈0.5 CU/ml for 20 min, and aggregation is insensitive to the presence of antagonists such as prostaglandin (PG) E1 and apyrase. Plasmin-induced platelet aggregability is associated with the expression of active fibrinogen receptors on the cell surface, which, using a l25I-fibrinogen binding assay, can be quantified to ≈2,300 molecules per platelet. Exposure of active αIIbβ3 receptors appears to depend partially, but not totally on a metabolic activation and granule exocytosis at the time of incubation with plasmin. In contrast with a-thrombin, plasmin-induced activation of αIIbβ3 is sustained and cannot be reversed by exposure of platelets to PGE1. Immunoblotting analysis of the receptor subunits shows no extensive proteolytic modification of αIIbβ3 by plasmin, and only reveals a limited proteolysis of the aminoterminal domain of the αIIb subunit. In addition to their capacity to aggregate in the presence of fibrinogen alone, plasmin-treated platelets also show a potentiated aggregability in response to low doses of ADP. Thus, plasmin has the potential to activate the platelet fibrinogen receptor in such a way that it remains irreversibly available to fibrinogen on the surface of nonaggregated cells, a feature that may participate to pathological states of in vivo platelet hyperaggregability.
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References
- 1 Anglés-Cano E. Overview on fibrinolysis: plasminogen activation pathways on fibrin and cell surfaces. Chem Phys Lipids 1994; 67/68: 353-362
- 2 Robbins KC. Fibrinolytic therapy: biochemical mechanisms. Semin Thromb Hemostas 1991; 17: 1-6
- 3 Collen D, Lijnen HR. Basic and clinical aspects of fibrinolysis and thrombolysis. Blood 1991; 78: 3114-3124
- 4 Coller BS. Platelets and thrombolytic therapy. N Engl J Med 1990; 322: 33-42
- 5 Fitzgerald DJ, Catella F, Roy L, FitzGerald GA. Marked platelet activation in vivo after intravenous streptokinase in patients with acute myocardial infarction. Circulation 1988; 77: 142-150
- 6 Hoffmann JJ M L, Bonnier JJ R M, Melman PG, Bartholomeus I, Couvée JE, Standring R, Fears R. Platelet function during thrombolytic therapy with anistreplase. Fibrinolysis 1993; 7: 401-407
- 7 Bertolino G, Noris P, Previtali M, Gamba G, Ferrario M, Montani N, Balduini CL. Platelet function after in vivo and in vitro treatment with thrombolytic agents. Am J Cardiol 1992; 69: 457-461
- 8 Torr SR, Winters KJ, Santoro SA, Sobel BE. The nature of interactions between tissue-type plasminogen activator and platelets. Thromb Res 1990; 59: 279-293
- 9 Gouin I, Lecompte T, Morel M-C, Lebrazi J, Modderman PW, Kaplan C, Samama MM. In vitro effects of plasmin on human platelet function in plasma. Inhibition of aggregation caused by fibrinogenolysis Circulation 1992; 85: 935-941
- 10 Parise P, Hauert J, Iorio A, Callegari P, Nenci GG. Fibrinogen degradation products generation is the major determinant of platelet inhibition induced by plasminogen activators in platelet-rich plasma. Fibrinolysis 1993; 7: 379-385
- 11 Schafer Al, Maas AK, Ware JA, Johnson PC, Rittenhouse SE, Salzman EW. Platelet protein phosphorylation, elevation of cytosolic calcium, and inositol phospholipid breakdown in platelet activation induced by plasmin. J Clin Invest 1986; 78: 73-79
- 12 Cramer EM, Lu H, Caen JP, Soria C, Bemdt MC, Tenza D. Differential redistribution of platelet glycoproteins Ib and IIb-IIIa after plasmin stimulation. Blood 1991; 77: 694-699
- 13 Lu H, Soria C, Cramer EM, Soria J, Maclouf J, Perrot JY, Li H, Commin PL, Schumann F, Regnier O, Lijnen RH, Caen JP. Temperature dependence of plasmin-induced activation or inhibition of human platelets. Blood 1991; 77: 996-1005
- 14 Penny WF, Ware JA. Platelet activation and subsequent inhibition by plasmin and recombinant tissue-type plasminogen activator. Blood 1992; 79: 91-98
- 15 Michelson AD, Barnard MR. Plasmin-induced redistribution of platelet glycoprotein Ib. Blood 1990; 76: 2005-2010
- 16 Lu H, Soria C, Soria J, De Romeuf C, Perrot J-Y, Tenza D, Garcia I, Caen JP, Martin-Cramer E. Reversible translocation of glycoprotein Ib in plas- min-treated platelets: consequences for platelet function. Eur J Clin Invest 1993; 23: 785-793
- 17 Schafer AI, Adelman B. Plasmin inhibition of platelet function and of arachidonic acid metabolism. J Clin Invest 1985; 75: 456-461
- 18 Niewiarowski S, Senyi AF, Gillies P. Plasmin-induced platelet aggregation and platelet release reaction. J Clin Invest 1973; 52: 1647-1659
- 19 Lopez JA. The platelet glycoprotein Ib-IX complex. Blood Coagul Fibrinol 1994; 5: 97-119
- 20 Strieker RB, Wong D, Shiu DT, Reyes PT, Shuman MA. Activation of plasminogen by tissue plasminogen activator on normal and thrombasthenic platelets: effects on surface proteins and platelet aggregation. Blood 1986; 68: 275-280
- 21 Adelman B, Michelson AD, Loscalzo J, Greenberg J, Handin RI. Plasmin effect on platelet glycoprotein Ib-von Willebrand factor interactions. Blood 1985; 65: 32-40
- 22 Pintigny D, Bihour C, Hourdillé P, Nurden AT. Multiple effects of plasmin on platelets. Thromb Haemost 1989; 62: 506 (abstr.)
- 23 Smyth SS, Joneckis CC, Parise LV. Regulation of vascular integrins. Blood 1993; 81: 2827-2843
- 24 Calvete JJ. Elements for a structural/functional model of human platelet plasma membrane fibrinogen receptor, the glycoprotein IIb/IIIa complex (integrin ±IIb²3). In: Cell Adhesion Molecules. Hemler ME, Mihich E. (eds) New York, NY: Plenum Press; 1993. pp 63-91
- 25 Peerschke EI B, Coller BS. A murine monoclonal antibody that blocks fibrinogen binding to normal platelets also inhibits fibrinogen interactions with chymotrypsin-treated platelets. Blood 1984; 64: 59-63
- 26 Pidard D, Frelinger AL, Bouillot C, Nurden AT. Activation of the fibrinogen receptor on human platelets exposed to alpha-chymotrypsin. Relationship with proteolytic cleavage at the carboxyterminus of the membrane glycoprotein IIb heavy chain Eur J Biochem 1991; 200: 437-447
- 27 Pidard D, Théodet J-C, Renesto P, Chignard M. The effects of serine proteinases on the platelet fibrinogen receptor. Thromb Haemost 1991; 65: 795 (abstr)
- 28 Lu H, Soria C, Li H, Soria J, Lijnen RH, Perrot JY, Caen JP. Role of active center and lysine binding sites of plasmin in plasmin-induced platelet activation and disaggregation. Thromb Haemost 1991; 65: 67-72
- 29 Bemdt MC, Du X, Booth WJ. Ristocetin-induced reconstitution of binding of von Willebrand factor to purified human platelet membrane glycoprotein Ib-IX complex. Biochemistry 1988; 27: 633-640
- 30 Pidard D, Montgomery RR, Bennett JS, Kunicki TJ. Interaction of AP-2, a monoclonal antibody specific for the human platelet glycoprotein IIb-IIIa complex, with intact platelets. J Biol Chem 1983; 258: 12582-12586
- 31 Legrand C, Dubemard V, Nurden AT. Studies on the mechanism of expression of secreted fibrinogen on the surface of activated human platelets. Blood 1989; 73: 1226-1234
- 32 Nurden AT, Dupuis D, Kunicki TJ, Caen JP. Analysis of the glycoprotein and protein composition of Bemard-Soulier platelets by single and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J Clin Invest 1981; 67: 1431-1440
- 33 Marguerie GA, Ginsberg MH, Plow EF. The platelet fibrinogen receptor. In: Platelets in Biology and Pathology III. Macintyre DE, Gordon JL. (eds) Amsterdam: Elsevier Science Publishers BV; 1987. pp 95-125
- 34 Legrand C, Dubemard V, Nurden AT. Characteristics of collagen-induced fibrinogen binding to human platelets. Biochim Biophys Acta 1985; 812: 802-810
- 35 Miller JL, Katz AJ, Feinstein MB. Plasmin inhibition of thrombin-induced platelet aggregation. Thromb Diathes Haemorrh 1975; 33: 286-309
- 36 Loscalzo J, Vaughan DE. Tissue plasminogen activator promotes platelet disaggregation in plasma. J Clin Invest 1987; 79: 1749-1755
- 37 Winters KJ, Eisenberg PR, Jaffe AS, Santoro SA. Dependence of plasminmediated degradation of platelet adhesive receptors on temperature and Ca2+. Blood 1990; 76: 1546-1557
- 38 Hoffmann JJ M L, Janssen WC M. Interactions between thrombolytic agents and platelets: effects of plasmin on platelet glycoproteins Ib and IIb/IIIa. Thromb Res 1992; 67: 711-719
- 39 Pasche B, Ouimet H, Francis S, Loscalzo J. Structural changes in platelet glycoprotein IIb/IIIa by plasmin: determinants and functional consequences. Blood 1994; 83: 404-414
- 40 Van Willigen G, Akkerman J-WN. Regulation of glycoprotein IIB/IIIA exposure on platelets stimulated with a-thrombin. Blood 1992; 79: 82-90