PDF herunterladen
Thromb Haemost 1994; 71(03): 347-352
DOI: 10.1055/s-0038-1642441
Original Article
Schattauer GmbH Stuttgart

Platelet-Bound Prekallikrein Promotes Pro-Urokinase-Induced Clot Lysis: A Mechanism for Targeting the Factor XII Dependent Intrinsic Pathway of Fibrinolysis

Jean-Pierre Loza
The Vascular Research Laboratory, Department of Medicine, Institute for the Prevention of Cardiovascular Disease, Deaconess Hospital, Harvard Medical School, Boston, MA, USA
,
Victor Gurewich
The Vascular Research Laboratory, Department of Medicine, Institute for the Prevention of Cardiovascular Disease, Deaconess Hospital, Harvard Medical School, Boston, MA, USA
,
Michael Johnstone
The Vascular Research Laboratory, Department of Medicine, Institute for the Prevention of Cardiovascular Disease, Deaconess Hospital, Harvard Medical School, Boston, MA, USA
,
Ralph Pannell
The Vascular Research Laboratory, Department of Medicine, Institute for the Prevention of Cardiovascular Disease, Deaconess Hospital, Harvard Medical School, Boston, MA, USA
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received: 17. Juni 1993

Accepted after revision 10. November 1993

Publikationsdatum:
06. Juli 2018 (online)

  als PDF herunterladen  Lizenzen und Reprints

Summary

Clots formed from platelet rich plasma were found to be lysed more readily by low concentrations of pro-urokinase (pro-UK) than clots formed from platelet poor plasma. This was not a non-specific effect since the reverse occurred with tissue plasminogen activator. A mechanical explanation due to platelet-mediated clot retraction was excluded by experiments in which retraction was inhibited with cyto-chalasin B. Therefore, a platelet-mediated enzymatic mechanism was postulated to explain the promotion of fibrinolysis. Casein autography of isolated platelets revealed a ≈ 90 kDa band of activity which comigrated with plasma prekallikrein (PK)/kallikrein, a known activator of pro-UK. Furthermore, treatment of platelets with plasma PK activator (PPA), consisting essentially of factor XIIa, induced activation of pro-UK and of chromomgenic substrate for kallikrein (S-2302). This activity corresponded to approximately 40-200 pM kallikrein per 10 8 washed and gel filtered platelets per ml. The activation of pro-UK by PPA-pretreated platelets was dose-dependent and inhibited by soybean trypsin inhibitor but not by bdellin, a specific inhibitor of plasmin, nor by the corn inhibitor of factor XIIa. Kinetic analysis of pro-UK activation by kallikrein showed promotion of the reaction by platelets. The KM of the reaction was reduced by platelets by ≈ 7-fold, while the kcat was essentially unchanged. In conclusion, PK was shown to be tightly associated with platelets where it can be activated by factor XIIa during clotting. The activation of pro-UK by platelet-bound kallikrein provides an explanation for the observed platelet mediated promotion of pro-UK-induced clot lysis. Since pro-UK and plasminogen have also been shown to be associated with platelets, the present findings suggest a mechanism by which the factor Xlla-dependent intrinsic pathway of fibrinolysis can be localized and targeted to a thrombus.

 

  • References

  • 1 Joist HJ. Platelets and fibrinolysis. Thrombos Haemostas 1977; 38: 955-62
    PubMedGoogle Scholar
  • 2 Johnson SA, Schneider CL. The existence of antifibrinolytic activity in platelets. Science 1953; 117: 229-32
    CrossrefPubMedGoogle Scholar
  • 3 Plow EF, Collen D. The presence and release of α2-antiplasmin from platelets. Blood 1981; 58: 1069-74
    PubMedGoogle Scholar
  • 4 Erickson LA, Ginsberg MH, Loskitoff DL. Detection and partial characterization of an inhibitor of plasminogen activator in human platelets. J Clin Invest 1984; 74: 1465-72
    CrossrefPubMedGoogle Scholar
  • 5 Kunitada S, Fitzgerald GA, Fitzgerald DJ. Inhibition of clot lysis and decreased binding of tissue plasminogen activator as a consequence of clot retraction. Blood 1992; 70: 1420-7
    PubMedGoogle Scholar
  • 6 Reed GL, Mastsueda GR, Haber E. Platelet factor XIII increases the fibrinolytic resistance of platelet-rich clots by accelerating the crosslinking of arantiplasmin to fibrin. Thrombos Haemostas 1992; 68: 315-20
    PubMedGoogle Scholar
  • 7 Miles LA, Plow EF. Binding and activation of plasminogen on the platelet surface. J Biol Chem 1985; 206: 4303-11
    PubMedGoogle Scholar
  • 8 Vaughn DE, Mendelsohn ME, De Clerck PJ, Collen D. Characterization of the binding of human tissue-type plasminogen activator to platelets. J Biol Chem 1989; 264: 15869-74
    PubMedGoogle Scholar
  • 9 Deguchi K, Shirakwa S. Plasminogen activation by tissue plasminogen activator in the presence of platelets. Thromb Res 1988; Suppl VIII: 65-72
    PubMedGoogle Scholar
  • 10 Gao SW, Morser J, MCLean K, Shuman M. Differential effect of platelets on plasminogen activation by tissue plasminogen activator, urokinase and streptokinase. Thromb Res 1990; 58: 421-33
    CrossrefPubMedGoogle Scholar
  • 11 Gurewich V, Pannell R. A comparative study of the efficacy and specificity of tissue plasminogen activator and pro-urokinase: demonstration of synergism and of different thresholds of nonselectivity. Thromb Res 1986; 44: 217-28
    CrossrefPubMedGoogle Scholar
  • 12 Gurewich V, Emmons F, Pannell R. Spontaneous clot lysis in whole human plasma by endogenous tissue type and urokinase type plasminogen activators: demonstration of a promoting effect by t-PA and by platelets on urokinase. Fibrinolysis 1988; 2: 143-9
    PubMedGoogle Scholar
  • 13 Gurewich V, Johnstone M, Loza JP, Pannell R. Prourokinase and pre- kallikrein are both associated with platelets: Implications for the intrinsic pathway of fibrinolysis and for therapeutic thrombolysis. FEBS Lett 1993; 318: 317-21
    CrossrefPubMedGoogle Scholar
  • 14 Mandle Jr RJ, Colman RW, Kaplan AP. Identification of prekallikrein and high-molecular-weight kininogen as a complex in human plasma. Proc Nat Acad Sci USA 1976; 73: 4179-83
    CrossrefPubMedGoogle Scholar
  • 15 Greengard JS, Griffin JH. Receptors for high molecular weight kininogen on stimulated washed human platelets. Biochemistry 1984; 23: 6863-9
    CrossrefPubMedGoogle Scholar
  • 16 Schmaier AH, Smith PA, Purdon AD, White JG, Colman W. High molecular weight kininogen: localization in the unstimulated and activated platelet and activation by a platelet calpain(s). Blood 1986; 67: 119-30
    PubMedGoogle Scholar
  • 17 Meloni FJ, Gustafson EJ, Schmaier AH. High molecular weight kininogen binds to platelets by its heavy and light chains and when bound has altered susceptibility to kallikrein cleavage. Blood 1992; 79: 1233-44
    CrossrefPubMedGoogle Scholar
  • 18 Ichinose A, Fujikawa K, Suyama T. The activation of prourokinase by plasma kallikrein and its inactivation by thrombin. J Biol Chem 1986; 261: 3486-9
    PubMedGoogle Scholar
  • 19 Hojima Y, Pierce JV, Pisano JJ. Hageman factor fragment inhibitor in corn seeds: purificaton and characterization. Thromb Res 1980; 20: 149-62
    CrossrefPubMedGoogle Scholar
  • 20 Orsini G, Brandazza A, Sarmientos P, Molinari A, Lansen J, Cauet G. Efficient renaturation and fibrinolytic properties of prourokinase and a deletion mutant expressed in Escherichia coli as inclusion bodies. Eur J Biochem 1991; 195: 691-7
    CrossrefPubMedGoogle Scholar
  • 21 Pannell R, Gurewich V. The activation of plasminogen by single-chain urokinase or by two-chain urokinase: a demonstration that single-chain has a low catalytic activity (prourokinase). Blood 1987; 69: 22-6
    PubMedGoogle Scholar
  • 22 Castellino FJ, Powell JR. Human plasminogen. Methods Enzymol 1981; 80: 365-78
    PubMedGoogle Scholar
  • 23 Mustard JF, Perry DW, Ardlie NG, Packham MA. Preparation of suspension of washed platelets from humans. Brit J Haemat 1972; 22: 193-204
    CrossrefPubMedGoogle Scholar
  • 24 Gurewich V, Pannell R, Louie S, Kelley P, Suddith RL, Greenlee R. Effective and fibrin-specific clot lysis by a zymogen precursor form of urokinase (pro-urokinase) A study in vitro and in two animal species. J Clin Invest 1984; 73: 1731-9
    CrossrefPubMedGoogle Scholar
  • 25 Fox JEB, Phillips DR. lnhibiton of actin polymerization in blood platelets by cytochalasins. Nature 1981; 292: 650-1
    CrossrefPubMedGoogle Scholar
  • 26 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 1970; 227: 680-5
    CrossrefPubMedGoogle Scholar
  • 27 Granelli-Piperno A, Reich E. A study of proteases and protease-inhibitor complexes in biological fluids. J Exp Med 1978; 148: 223-34
    CrossrefPubMedGoogle Scholar
  • 28 Vassalli JD, Dayer JM, Wohlwend A, Belin D. Concomitant secretion of prourokinase and of a plasminogen activator specific inhibitor by cultured human monocytes macrophages. J Exp Med 1984; 159: 1653-68
    CrossrefPubMedGoogle Scholar
  • 29 Pannell R, Gurewich V. Direct analysis of plasminogen activator species in whole plasma. Fibrinolysis 1992; 6 (Suppl 2) 128
    PubMedGoogle Scholar
  • 30 Clubber BAK, Morris JP, Castellino FJ. Effects of human fibrinogen and its cleavage products on activation of human plasminogen by streptokinase. Biochemistry 1985; 24: 3429-34
    CrossrefPubMedGoogle Scholar
  • 31 Liu Jn, Gurewich V. Fragment E-2 from fibrin substantially enhances prourokinase-induced Glu-plasminogen activation. A kinetic study using the plasmin-resistant mutant pro-urokinase Ala-158-rpro-UK. Biochem 1992; 31: 6311-7
    CrossrefPubMedGoogle Scholar
  • 32 Bradford MM. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-54
    CrossrefPubMedGoogle Scholar
  • 33 Nagase H, Barnett AJ, Human plasma kallikrein. A rapid purification method with high yield. Biochem J 1981; 193: 187-92
    CrossrefPubMedGoogle Scholar
  • 34 Fritz H, Krejci K. Trypsin-plasmin inhibitors (Bdellins) from leeches. Meth Enzymol 1976; 45: 797-806
    PubMedGoogle Scholar
  • 35 Gurewich V, Pannell R, Broeze RJ, Mao JI. Characterization of the intrinsic fibrinolytic properties of pro-urokinase through a study of plasmin- resistant mutant forms produced by site-specific mutagenesis of lysine-158. J Clin Invest 1988; 82: 1956-62
    CrossrefPubMedGoogle Scholar
  • 36 De Clerck PJ, Lijnen HR, Verstreken M, Moreau H, Collen D. A monoclonal antibody specific for two-chain urokinase-type plasminogen activator. Application to the study of the mechanism of clot lysis with single-chain urokinase- type plasminogen activator in plasma. Blood 1990; 75: 1794-1800
    PubMedGoogle Scholar
  • 37 Park S, Harker LA, Marzec UM, Levin EG. Demonstration of single chain urokinase-type plasminogen activator on human platelet membrane. Blood 1989; 73: 1421-5
    PubMedGoogle Scholar
  • 38 Wun TC, Capuano A. Spontaneous fibrinolysis in whole plasma. Identification of tissue activator as the major plasminogen activator causing spontaneous activity in vitro. J Biol Chem 1985; 260: 5061-6
    PubMedGoogle Scholar
  • 39 Hauert J, Nicoloso G, Schleuning WD, Bachmann F, Schapira M. Plasminogen activators in dextran sulfate activated euglobulin fractions. A molecular analysis of factor XII and prekallikrein-dependent fibrinolysis. Blood 1989; 73: 994-9
    PubMedGoogle Scholar
  • 40 Colman RW. Surface-mediated defense reactions. The plasma contact activation system. J Clin Invest 198-1 73: 1249-53
    PubMedGoogle Scholar
  • 41 Nievviarowski S, Prou-Wartelle O. Role du faeleui contact (faclcur Hageman) dans la fibrinolyse. Thromb Diath Haemorrh 1959; 3: 593-603
    PubMedGoogle Scholar
  • 42 Kluft C, Donijewaard G, Emeis JJ. Role of the contact system in fibrinoly sis. Semin Thromb Hemostas 1987; 13: 50-68
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 43 Ratnoff OD. A quarter century with Mr Hageman. Thromb Hacmostas 1980; 43: 95-8
    PubMedGoogle Scholar
  • 44 Goodnough LT, Saito H, Ratnoff OD. Thrombosis or myocardial infarction in congenital clotting factor abnormalities and chronic thrombocytopenias: A report of 21 patients and a review of 50 previously reported cases. Medicine (Baltimore) 1983; 62: 218-55
    PubMedGoogle Scholar
  • 45 Saito H. Contact factors in health and disease. Sem Thromb Hemostas 1987; 13: 36-49
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 46 Miles LA, Greengard JS, Griffin JH. A comparison of the abilities of plasma kallikrein, (3-factor Xlla, factor XIa and urokinase to activate plasminogen. Thromb Res 1983; 29: 407-17
    CrossrefPubMedGoogle Scholar
  • 47 Tsuda H, Miyata T, Iwanaga S, Yarnamoto T. Analysis of intrinsic fibrinolysis in human plasma induced by dextran sulfate. Thromb Haemostas 1992; 67: 440-4
    PubMedGoogle Scholar