Thromb Haemost 1998; 80(01): 161-166
DOI: 10.1055/s-0037-1615156
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Human Thrombin and Calcium Bound Factor Xa Significantly Shorten tPA-Induced Fibrin Clot Lysis Time via Neutralization of Plasminogen Activator Inhibitor Type 1 Activity

Tetsumei Urano
1   Departments of Physiology and Pathophysiology
,
Nobuo Nagai
1   Departments of Physiology and Pathophysiology
,
Megumi Matsuura
1   Departments of Physiology and Pathophysiology
,
Hayato Ihara
1   Departments of Physiology and Pathophysiology
,
Yumiko Takada
2   Hamamatsu University, School of Medicine, Hamamatsu, Shizuoka, Japan
,
Akikazu Takada
1   Departments of Physiology and Pathophysiology
› Author Affiliations
Further Information

Publication History

Received 30 November 1997

Accepted after revision 26 February 1998

Publication Date:
08 December 2017 (online)

Summary

Employing a fibrin clot lysis assay, we reassessed the significance of the neutralization of plasminogen activator inhibitor type 1 (PAI-1) activity by thrombin and factor-Xa in fibrinolysis. When PAI-1 enriched fibrin clots were formed using increasing concentrations of thrombin (0.1, 0.5, 1.0 IU/ml), their lysis times became shorter (43.8 ± 4.9, 25.7 ± 3.7, 13.9 ± 0.8 h respectively). Times were shortened further by either heparin (43.9 ± 11.0, 12.1 ± 2.6, 3.6 ± 0.2 h respectively) or vitronectin (17.0 ± 1.6, 1.9 ± 0.7, 0.9 ± 0.0 h respectively). Factor-Xa together with Ca++ shortened the clot lysis time further. Fibrin autography revealed that both enzymes dose dependently interfered with complex formation between tPA and PAI-1, making large amounts of tPA remaining to be free form. The mechanism seems to play a role in the coagulation associated enhancement of fibrinolysis.

 
  • References

  • 1 Loskutoff DJ, Sawdey M, Mimuro J. Type 1 plasminogen activator inhibitor. Prog Hemost Thromb 1989; 9: 87-115.
  • 2 Castellino FJ. Recent advances in the chemistry of the fibrinolytic system. Chem Rev 1981; 81: 431-46.
  • 3 Collen D. On the regulation and control of fibrinolysis. Edward Kowalski Memorial Lecture. Thromb Haemost 1980; 43: 77-89.
  • 4 Carrell RW, Evans DL, Stein PE. Mobile reactive centre of serpins and the control of thrombosis. Nature 1991; 353: 576-8.
  • 5 Doolittle RF. Angiotensinogen is related to the antitrypsin-antithrombinovalbumin family. Science 1983; 222: 417-9.
  • 6 Travis J, Salvesen GS. Human plasma proteinase inhibitors. Annu Rev Biochem 1983; 52: 655-709.
  • 7 Huber R, Carrell RW. Implications of the three-dimensional structure of alpha 1-antitrypsin for structure and function of serpins. Biochemistry 1989; 28: 8951-66.
  • 8 Edelberg JM, Reilly CF, Pizzo SV. The inhibition of tissue type plasminogen activator by plasminogen activator inhibitor-1. The effects of fibrinogen, heparin, vitronectin, and lipoprotein(a). J Biol Chem 1991; 266: 7488-93.
  • 9 Urano T, Serizawa K, Takada Y, Ny T, Takada A. Heparin and heparan sul-fate enhancement of the inhibitory activity of plasminogen activator inhibitor type 1 toward urokinase type plasminogen activator. Biochim Biophys Acta 1994; 1201: 217-22.
  • 10 Ehrlich HJ, Gebbink RK, Keijer J, Linders M, Preissner KT, Pannekoek H. Alteration of serpin specificity by a protein cofactor. Vitronectin endows plasminogen activator inhibitor 1 with thrombin inhibitory properties. J Biol Chem 1990; 265: 13029-35.
  • 11 Keijer J, Linders M, Wegman JJ, Ehrlich HJ, Mertens K, Pannekoek H. On the target specificity of plasminogen activator inhibitor-1: the role of heparin, vitronectin, and the reactive site. Blood 1991; 78: 1254-61.
  • 12 Urano T, Ihara H, Takada Y, Nagai N, Takada A. The inhibition of human factor Xa by plasminogen activator inhibitor type 1 in the presence of calcium ion, and its enhancement by heparin and vitronectin. Biochim Biophys Acta 1996; 1298: 199-208.
  • 13 Fa M, Karolin J, Aleshkov S, Strandberg L, Johansson LB, Ny T. Time-resolved polarized fluorescence spectroscopy studies of plasminogen activator inhibitor type 1: conformational changes of the reactive center upon interactions with target proteases, vitronectin and heparin. Biochemistry 1995; 34: 13833-40.
  • 14 Ehrlich HJ, Gebbink RK, Preissner KT, Keijer J, Esmon NL, Mertens K, Pannekoek H. Thrombin neutralizes plasminogen activator inhibitor-1 (PAI-1) that is complexed with vitronectin in the endothelial cell matrix. J Cell Biol 1991; 115: 1773-81.
  • 15 Urano T, Strandberg L, Johansson LB, Ny T. A substrate-like form of plasminogen-activator-inhibitor type 1. Conversions between different forms by sodium dodecyl sulphate. Eur J Biochem 1992; 209: 985-92.
  • 16 Lawrence D, Strandberg L, Grundström T, Ny T. Purification of active hu man plasminogen activator inhibitor 1 from Escherichia coli. Comparison with natural and recombinant forms purified from eucaryotic cells. Eur J Biochem 1989; 186: 523-33.
  • 17 Jameson GW, Roberts DV, Adams RW, Kyle WS, Elmore DT. Determination of the operational molarity of solutions of bovine alpha-chymotrypsin, trypsin, thrombin and factor Xa by spectrofluorimetric titration. Biochem J 1973; 131: 107-17.
  • 18 Deutsch DG, Mertz ET. Plasminogen: purification from human plasma by affinity chromatography. Science 1970; 170: 1095-6.
  • 19 Urano S, Metzger AR, Castellino FJ. Plasmin-mediated fibrinolysis by variant recombinant tissue plasminogen activators. Proc Natl Acad Sci USA 1989; 86: 2568-71.
  • 20 Carlson RH, Garnick RL, Jones AJ, Meunier AM. The determination of recombinant human tissue-type plasminogen activator activity by turbidimetry using a microcentrifugal analyzer. Anal Biochem 1988; 168: 428-35.
  • 21 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5.
  • 22 Granelli PA, Reich E. A study of proteases and protease-inhibitor complexes in biological fluids. J Exp Med 1978; 148: 223-34.
  • 23 Patston PA, Schapira M. Low-affinity heparin stimulates the inactivation of plasminogen activator inhibitor-1 by thrombin. Blood 1994; 84: 1164-72.
  • 24 de Fouw Nj, van Hinsbergh VW, de Jong Yf, Haverkate F, Bertina RM. The interaction of activated protein C and thrombin with the plasminogen activator inhibitor released from human endothelial cells. Thromb Haemost 1987; 57: 176-82.
  • 25 Madden RM, Levin EG, Marlar RA. Thrombin and the thrombin-thrombomodulin complex interaction with plasminogen activator inhibitor type-1. Blood Coagul Fibrinolysis 1991; 2: 471-6.
  • 26 Naski MC, Lawrence DA, Mosher DF, Podor TJ, Ginsburg D. Kinetics of inactivation of alpha-thrombin by plasminogen activator inhibitor-1. Comparison of the effects of native and urea-treated forms of vitronectin. J Biol Chem 1993; 268: 12367-72.
  • 27 Gebbink RK, Reynolds CH, Tollefsen DM, Mertens K, Pannekoek H. Specific glycosaminoglycans support the inhibition of thrombin by plasminogen activator inhibitor-1. Biochemistry 1993; 32: 1675-80.
  • 28 Gabriel DA, Muga K, Boothroyd EM. The effect of fibrin structure on fibrinolysis. J Biol Chem 1992; 267: 24259-63.
  • 29 Jackson CM. Factor X. Prog Hemost Thromb 1984; 7: 55-109.
  • 30 Urano T, Sumiyoshi K, Pietraszek MH, Takada Y, Takada A. PAI-1 plays an important role in the expression of t-PA activity in the euglobulin clot lysis by controlling the concentration of free t-PA. Thromb Haemost 1991; 66: 474-8.
  • 31 Urano T, Sakakibara K, Rydzewski A, Urano S, Takada Y, Takada A. Relationships between euglobulin clot lysis time and the plasma levels of tissue plasminogen activator and plasminogen activator inhibitor-1. Thromb Haemost 1990; 63: 82-6.
  • 32 Kojima Y, Urano T, Kojima K, Serizawa K, Takada Y, Takada A. The significant enhancement of fibrinolysis by calcium ion in a cell free system: the shortening of euglobulin clot lysis time by calcium ion. Thromb Haemost 1994; 72: 113-8.
  • 33 Urano T, Chibber BA, Castellino FJ. The reciprocal effects of epsilon-aminohexanoic acid and chloride ion on the activation of human [Glu1]plasminogen by human urokinase. Proc Natl Acad Sci USA 1987; 84: 4031-4.
  • 34 Hoylaerts M, Rijken DC, Lijnen HR, Collen D. Kinetics of the activation of plasminogen by human tissue plasminogen activator. Role of fibrin. J Biol Chem 1982; 257: 2912-9.
  • 35 Sakata Y, Aoki N. Cross-linking of alpha 2-plasmin inhibitor to fibrin by fibrin-stabilizing factor. J Clin Invest 1980; 65: 290-7.
  • 36 Wu K, Urano T, Ihara H, Takada Y, Fujie M, Shikimori M, Hashimoto K, Takada A. The cleavage and inactivation of plasminogen activator inhibitor type 1 by neutrophil elastase: The evaluation of its physiologic relevance in fibrinolysis. Blood 1995; 86: 1056-61.
  • 37 Bajzar L, Morser J, Nesheim M. TAFI, or plasma procarboxypeptidase B, couples the coagulation and fibrinolytic cascades through the thrombinthrombomodulin complex. J Biol Chem 1996; 271: 16603-8.
  • 38 Bajzar L, Manuel R, Nesheim ME. Purification and characterization of TAFI, a thrombin-activable fibrinolysis inhibitor. J Biol Chem 1995; 270: 14477-84.