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

Jean-Pierre Loza; Victor Gurewich; Michael Johnstone; Ralph Pannell

doi:10.1055/s-0038-1642441

Thrombosis and Haemostasis, Table of Contents

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

› Author Affiliations

Abstract

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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 XII_a, 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 K_M of the reaction was reduced by platelets by ≈ 7-fold, while the k_cat 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.

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References

References
1 Joist HJ. Platelets and fibrinolysis. Thrombos Haemostas 1977; 38: 955-62
2 Johnson SA, Schneider CL. The existence of antifibrinolytic activity in platelets. Science 1953; 117: 229-32
3 Plow EF, Collen D. The presence and release of α₂-antiplasmin from platelets. Blood 1981; 58: 1069-74
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
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
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
7 Miles LA, Plow EF. Binding and activation of plasminogen on the platelet surface. J Biol Chem 1985; 206: 4303-11
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
9 Deguchi K, Shirakwa S. Plasminogen activation by tissue plasminogen activator in the presence of platelets. Thromb Res 1988; Suppl VIII: 65-72
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
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
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
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
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
15 Greengard JS, Griffin JH. Receptors for high molecular weight kininogen on stimulated washed human platelets. Biochemistry 1984; 23: 6863-9
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
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
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
19 Hojima Y, Pierce JV, Pisano JJ. Hageman factor fragment inhibitor in corn seeds: purificaton and characterization. Thromb Res 1980; 20: 149-62
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
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
22 Castellino FJ, Powell JR. Human plasminogen. Methods Enzymol 1981; 80: 365-78
23 Mustard JF, Perry DW, Ardlie NG, Packham MA. Preparation of suspension of washed platelets from humans. Brit J Haemat 1972; 22: 193-204
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
25 Fox JEB, Phillips DR. lnhibiton of actin polymerization in blood platelets by cytochalasins. Nature 1981; 292: 650-1
26 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 1970; 227: 680-5
27 Granelli-Piperno A, Reich E. A study of proteases and protease-inhibitor complexes in biological fluids. J Exp Med 1978; 148: 223-34
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
29 Pannell R, Gurewich V. Direct analysis of plasminogen activator species in whole plasma. Fibrinolysis 1992; 6 (Suppl 2) 128
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
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
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
33 Nagase H, Barnett AJ, Human plasma kallikrein. A rapid purification method with high yield. Biochem J 1981; 193: 187-92
34 Fritz H, Krejci K. Trypsin-plasmin inhibitors (Bdellins) from leeches. Meth Enzymol 1976; 45: 797-806
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
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
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
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
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
40 Colman RW. Surface-mediated defense reactions. The plasma contact activation system. J Clin Invest 198-1 73: 1249-53
41 Nievviarowski S, Prou-Wartelle O. Role du faeleui contact (faclcur Hageman) dans la fibrinolyse. Thromb Diath Haemorrh 1959; 3: 593-603
42 Kluft C, Donijewaard G, Emeis JJ. Role of the contact system in fibrinoly sis. Semin Thromb Hemostas 1987; 13: 50-68
43 Ratnoff OD. A quarter century with Mr Hageman. Thromb Hacmostas 1980; 43: 95-8
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
45 Saito H. Contact factors in health and disease. Sem Thromb Hemostas 1987; 13: 36-49
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
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