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DOI: 10.1055/s-0038-1647456
Role of Active Center and Lysine Binding Sites of Plasmin in Plasmin-Induced Platelet Activation and Disaggregation
Publication History
Received 11 June 1990
Accepted after revision 03 August 1990
Publication Date:
04 September 2018 (online)
Summary
Previous studies have shown that plasmin can activate platelets and - can also disperse platelet aggregates by degradation of fibrinogen bound to platelets. In this study, the role of the active center and the lysine binding sites (LBS) of human plasmin in activating platelets and in dispersing platelet aggresates is investigated using aprotinin and the tripeptide Val-Phe-Lys-CH2Cl to inhibit the active center and using epsilon-aminocaproic acid (EACA) to specifically block the LBS. Our results show that the catalytic activity of plasmin is indispensable both for activating platelets and for dispersing platelet aggregates. Binding of plasmin to platelets through the LBS enhances its activating potential, since both EACA (1 mM) and Lys-plasminogen (molar ratio of plasminogen:plasmin at 2:l to 4:1) inhibit plasmininduced platelet activation, whereas Glu-plasminogen at a molar ratio of 15: I had no effect. Furthermore, plasmin which lacks the LBS (miniplasmin), is about 3 fold less effective in activating platelets. Flowever, plasmin binding through the LBS is not absolutely required to disperse platelet aggregates, since EACA at 30 mmol/l was unable to prevent disaggregation by plasmin (half disaggregation time: 40 min in the presence of EACA against 27 min in its absence). It also appeared that fibrinogen receptors on activated platelets are resistant to plasmin degradation, and that disaggregation of plasmin-induced platelet aggregates was much slower than the degradation of fibrinogen by plasmin.
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References
- 1 Niewiarowski S, Senyi AF, Gillies P. Plasmin-induced platelet activation and release reaction. Effect on hemostasis. J Clin Invest 1973; 52: 1647-1659
- 2 Miller JL, Katz AJ, Feinstein MB. Plasmin inhibition of thrombin-induced platelet aggregation. Thromb Diathes Haemorrh 1975; 33: 286-309
- 3 Schafer AI, Adelman B. Plasmin inhibition of platelet function and of arachidonic acid metabolism. J Clin Invest 1985; 75: 456-461
- 4 Guccione MA, Kinlough-Rathbone RL, Packham MA, Harfenist EJ, Rand ML, Greenberg JP, Perry DW, Mustard JF. Effects of plasmin on rabbit platelets. Thromb Haemostas 1985; 53: 8-14
- 5 Adnot S, Ferry N, Hanoune J, Lacombe ML. Plasmin: a possible physiological modulator of the human platelet adenylate cyclase system. Clin Sci 1987; 72: 467-473
- 6 Loscalzo J. Platelet and thrombolysis. J Vase Med Biol 1989; 1: 213-219
- 7 Coller BS. Platelets and thrombolytic therapy. N Engl J Med 1990; 322: 33-42
- 8 Schafer AI, Maas AN, 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
- 9 Greenberg J, Orr JL, Packman MA, Guccione MA, Harfenist EJ, Kinglough-Rathbone RL, Perry DW, Mustard JF. The effect of pretreatment of human or rabbit platelets with chymotrypsin on their responses to human fibrinogen and aggregating agents. Blood 1979; 54: 753-765
- 10 Niewiarowski S, Budzynski AZ, Morinelli TA, Brudzynnski TM, Stewart GJ. Exposure of fibrinogen receptor on human platelets by proteolytic enzymes. J Biol Chem 1979; 256: 917-925
- 11 Lu H, Soria C, Cramer EM, Soria J, Perrot JY, Li H, Commin PL, Schumann F, Regnier O, Lijnen HR, Caen JP. Temperature dependence of plasmin-induced activation or inhibition of human platelet. Blood (in press)
- 12 Rudd MA, Amarante P, Smick D, Vaughan DE, Loscalzo J. Temporal effects of tissue plasminogen activator infusion on platelet aggregation ex vivo. Blood 1988; 72 (Suppl. 01) 374A (Abstr)
- 13 Jang IK, Gold HK, Ziskind AA, Fallon JT, Holt RE, Leinbach RC, May JW, Collen D. Differential sensitivity of erythrocyte-rich and platelet-rich arterial thrombi to lysis with recombinant tissue-type plasminogen activator. A possible explanation for resistance to thrombolysis. Circulation 1989; 79: 920-928
- 14 Loscalzo J, Vaughan C. Tissue plasminogen activator promotes platelet disaggregation in plasma. J Clin Invest 1987; 79: 1749-1755
- 15 Castellino FJ. Biochemistry of human plasminogen. Semin Thromb Haemostas 1984; 10: 18-23
- 16 Lucas MA, Fretto LJ. The binding of plasminogen to fibrin and fibrinogen. J Biol Chem 1983; 258: 4249-4256
- 17 Miles LA, Plow EF. Binding and activation of plasminogen on the platelet surface. J Biol Chem 1985; 260: 4303-4311
- 18 Adelman B, Rizk A, Hanners E. Plasminogen interacts with platelets in plasma. Blood 1988; 72: 1530-1535
- 19 Adelman B, Michelson AD, Greenberg J, Handin R. Proteolysis of platelet glycoprotein Ib by plasmin is facilitated by plasmin lysine-binding regions. Blood 1986; 68: 1280-1284
- 20 Vaughan DE, Mendelsohn ME, Declerck PJ, Van Houtte EV, Loscalzo J. Characterization of the binding of human tissue-type plasminogen activator to platelets. J Biol Chem 1989; 264: 15869-15874
- 21 Park S, Harker LA, Marzec UM, Levin EG. Demonstration of single chain urokinase-type plasminogen activator on human platelet membrane. Blood 1989; 73: 1421-1425
- 22 Plow EF, Freaney DE, Plescia J, Miles LA. The plasminogen system and the cell surface: evidence for plasminogen and urokinase receptors on the same cell type. J Cell Biol 1986; 103: 2411-2420
- 23 Wiman B, Boman L, Collen D. On the kinetics of the reaction between human antiplasmin and a low molecular weight form of plasmin. Eur J Biochem 1978; 87: 143-146
- 24 Patscheke H, Womer P. Platelet activation detected by turbidimetric shape-change analysis. Thrombolysis Res 1978; 12: 485-496
- 25 Shattil SJ, Hoxie JA, Cunningham M, Brass LF. Changes in the platelet membrane glycoprotein Ilb/IIIa complex during platelet activation. J Biol Chem 1985; 260: 11107-11114
- 26 Larsen E, Celi A, Gibert GE, Furie BC, Erban JK, Bonfanti R, Wagner DD, Furie B. PADGEM protein: A receptor that mediates the interaction of activated platelets with neutrophils and monocytes. Cell 1989; 59: 305-312
- 27 Wiman B. On the reaction of plasmin or plasmin-streptokinase complex with aprotinin or α1-antiplasmin. Thromb Res 1980; 17: 143-152
- 28 Collen D, Lijnen HR, De Cock F, Durieux JP, Loffet A. Kinetic properties of tripeptide lysyl chloromethyl ketone and lysyl p-nitroanilide derivatives towards trypsin-like serine proteinase. Biochim Biophys Acta 1980; 165: 158-166
- 29 Okamoto S, Okamoto U, Okada Y, Hijikata-Okunomiya A, Wanaka K, Horie N, Naito T. Coding, decoding and noise of proteinase: a rational approach to plasmin inhibitors. In: Fundamental and Clinical Fibrinolysis Castellino FJ. (ed). Elsevier Science Publishers; Amsterdam: 1987: 67-82
- 30 Christensen U, Sottrup-Jensen L, Magnusson S, Petersen TE, Clemmensen J. Enzymatic properties of the neoplasmin-Val442 (mini-plasmin). Biochim Biophys Acta 1979; 567: 472-481
- 31 Sottrup-Jensen L, Claeys H, Lajdel M, Petersen TE, Magnusson S. The primary structure of human plasminogen Isolation of two lysine-binding fragments and one “mini-plasminogen” (Mw 38,000) by elastase catalysed specific limited proteolysis. In: Progress in Chemical Fibrinolysis and Thrombolysis Davidson JF, Roman RM, Samama MM. (eds). Raven Press; New York: 1978. 3 139-209
- 32 Caen JP, Larrieu MJ, Samama MM. L’Hemostase, méthodes d’exploration et de diagnostic pratique. L’Expansion scientifique francaise Paris: 1975
- 33 Marguierie G, Plow EF. The fibrinogen-dependent pathway of platelet aggregation. Ann NY Acad Sci 1983; 408: 556-566
- 34 Harfenist EJ, Packhan MA, Kinlough-Rathbone RL, Mustard JI. Inhibitors of ADP-induced platelet aggregation prevent fibrinogen binding to rabbit platelets and cause rapid deaggregation and dissociation of bound fibrinogen. J Lab Clin Med 1981; 97: 680-688
- 35 Peerschke EI B. Decreased accessibility of platelet-bound fibrinogen to antibody and enzyme probes. Blood 1989; 74: 682-689
- 36 Nachman RL, Silverstein RL, Asch AS. Thrombospondin: Cell biology of an adhesive glycoprotein. In: Thrombosis and Haemostasis Verstraete M, Vermylen J, Lijnen R, Arnout J. (eds). Leuven University Press; Leuven: 1987: 81-91
- 37 Parise LV, Phillips DR. Fibronectin-binding properties of the purified platelet glycoprotein Ilb/IIIa complex. J Biol Chem 1986; 261: 14011-14020
- 38 Plow EF, Sroujia AH, Meyer D, Marguerie G, Ginsberg MH. Evidence that three adhesive proteins interact with a common recognition site on activated platelets. J Biol Chem 1984; 259: 5388-5391
- 39 Kastrikina TF, Taran LD, Kudinov SA. Kinetic characteristics of fibrinogen and fibrin hydrolysis by plasmin 1 and 2 and miniplasmin. Thromb Res 1986; 41: 681-688
- 40 Morris JP, Castellino FG. The role of the lysine binding sites of human plasmin in the hydrolysis of human fibrinogen. Biochim Biophys Acta 1983; 744: 93-104