Download PDF
Thromb Haemost 2004; 91(03): 473-479
DOI: 10.1160/TH03-06-0377
Theme Issue Article
Schattauer GmbH

Thrombin activatable fibrinolysis inhibitor (TAFI) affects fibrinolysis in a plasminogen activator concentration-dependent manner

Study of seven plasminogen activators in an internal clot lysis model
Ana H. C. Guimarães
1   Gaubius Laboratory, TNO Prevention and Health, Leiden
2   the Department of Physiology, Institute for Cardiovascular Research, VU University, Amsterdam, The Netherlands
,
Dingeman C. Rijken
1   Gaubius Laboratory, TNO Prevention and Health, Leiden
3   the Department of Hematology, Erasmus University Medical Center, Rotterdam,The Netherlands
› Author Affiliations
Further Information

Publication History

Received 18 June 2003

Accepted after resubmission 26 February 2003

Publication Date:
27 December 2017 (online)

    Permissions and Reprints

Summary

TAFIa was shown to attenuate fibrinolysis. In our in vitro study, we investigated how the inhibitory effect of TAFIa depended on the type and concentration of the plasminogen activator (PA). We measured PA-mediated lysis times of plasma clots under conditions of maximal TAFI activation by thrombin-thrombomodulin in the absence and presence of potato carboxypeptidase inhibitor. Seven different PAs were compared comprising both tPA-related (tPA, TNK-tPA, DSPA), bacterial PA-related (staphylokinase and APSAC) and urokinase-related (tcu-PA and k2tu-PA) PAs. The lysis times and the retardation factor were plotted against the PA concentration. The retardation factor plots were bell-shaped. At low PA concentrations, the retardation factor was low, probably due to the limited stability of TAFIa. At intermediate PA concentrations the retardation factor was maximal (3-6 depending on the PA), with TNK-tPA, APSAC and DSPA exhibiting the strongest effect. At high PA concentrations, the retardation factor was again low, possibly due to inactivation of TAFIa by plasmin or to a complete conversion of glu-plasminogen into lys-plasminogen. Using individual plasmas with a reduced plasmin inhibitor activity (plasmin inhibitor Enschede) the bell-shaped curve of the retardation factor shifted towards lower tPA and DSPA concentrations, but the height did not decrease. In conclusion, TAFIa delays the lysis of plasma clots mediated by all the plasminogen activators tested. This delay is dependent on the type and concentration of the plasminogen activator, but not on the fibrin specificity of the plasminogen activator. Furthermore, plasmin inhibitor does not play a significant role in the inhibition of plasma clot lysis by TAFI.

Keywords

TAFI - internal lysis - plasminogen activator - plasmin inhibitor
 

  • References

  • 1 Nesheim M, Wang W, Boffa M. et al. Thrombin, thrombomodulin and TAFI in the molecular link between coagulation and fibrinolysis. Thromb Haemost 1997; 78: 386-91.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 2 Nesheim ME. TAFI. Fibrinolysis & Proteolysis 1999; 13: 72-77.
    CrossrefPubMedGoogle Scholar
  • 3 Bajzar L. Thrombin activatable fibrinolysis inhibitor and an antifibrinolytic pathway. Arterioscler Thromb Vasc Biol 2000; 20: 2511-8.
    CrossrefPubMedGoogle Scholar
  • 4 Bouma BN, Marx PF, Mosnier LO. et al. Thrombin-activatable fibrinolysis inhibitor (TAFI, plasma procarboxypeptidase B, procarboxypeptidase R, procarboxypeptidase U). Thromb Res 2001; 101: 329-354.
    CrossrefPubMedGoogle Scholar
  • 5 Bajzar L, Morser J, Nesheim M. TAFI, or plasma procarboxypeptidase B, couples the coagulation and fibrinolytic cascades through the thrombin-thrombomodulin complex. J Biol Chem 1996; 271: 16603-8.
    CrossrefPubMedGoogle Scholar
  • 6 Kokame K, Zheng X, Sadler JE. Activation of thrombin-activable fibrinolysis inhibitor requires epidermal growth factor-like domain 3 of thrombomodulin and is inhibited competitively by protein C. J Biol Chem 1998; 273: 12135-9.
    CrossrefPubMedGoogle Scholar
  • 7 Bajzar L, Nesheim M, Morser J. et al. Both cellular and soluble forms of thrombomodulin inhibit fibrinolysis by potentiating the activation of thrombin-activable fibrinolysis inhibitor. J Biol Chem 1998; 273: 2792-8.
    CrossrefPubMedGoogle Scholar
  • 8 Marx PF, Dawson PE, Bouma BN. et al. Plasmin-mediated activation and inactivation of thrombin-activatable fibrinolysis inhibitor. Biochemistry 2002; 41: 6688-96.
    CrossrefPubMedGoogle Scholar
  • 9 Redlitz A, Tan AK, Eaton DL, Plow EF. Plasma carboxypeptidases as regulators of the plasminogen system. J Clin Invest 1995; 96: 2534-8.
    CrossrefPubMedGoogle Scholar
  • 10 Broze GJJ, Higuchi DA. Coagulation-dependent inhibition of fibrinolysis: role of carboxypeptidase-U and the premature lysis of clots from hemophilic plasma. Blood 1996; 88: 3815-23.
    PubMedGoogle Scholar
  • 11 Sakharov DV, Plow EF, Rijken DC. On the mechanism of the antifibrinolytic activity of plasma carboxypeptidase B. J Biol Chem 1997; 272: 14477-82.
    CrossrefPubMedGoogle Scholar
  • 12 von dem Borne PA, Meijers JC, Nesheim ME. et al. Thrombin-mediated activation of factor XI results in a thrombin-activatable fibrinolysis inhibitor-dependent inhibition of fibrinolysis. J Clin Invest 1997; 99: 2323-7.
    CrossrefPubMedGoogle Scholar
  • 13 Wang W, Boffa MB, Bajzar L. et al. A study of the mechanism of inhibition of fibrinolysis by activated thrombin-activable fibrinolysis inhibitor. J Biol Chem 1998; 273: 27176-81.
    CrossrefPubMedGoogle Scholar
  • 14 Suenson E, Lutzen O, Thorsen S. Initial plasmin-degradation of fibrin as the basis of a positive feed-back mechanism in fibrinolysis. Eur J Biochem 1984; 140: 513-22.
    CrossrefPubMedGoogle Scholar
  • 15 Christensen U. C-terminal lysine residues of fibrinogen fragments essential for binding to plasminogen. FEBS Lett 1985; 182: 43-46.
    CrossrefPubMedGoogle Scholar
  • 16 Fleury V, Angles-Cano E. Characterization of the binding of plasminogen to fibrin surfaces: the role of carboxy-terminal lysines. Biochemistry 1991; 30: 7630-8.
    CrossrefPubMedGoogle Scholar
  • 17 Wiman B, Collen D. Purification and characterization of human antiplasmin, the fast-acting plasmin inhibitor in plasma. Eur J Biochem 1977; 78: 19-26.
    CrossrefPubMedGoogle Scholar
  • 18 Wiman B, Collen D. Molecular mechanism of physiological fibrinolysis. Nature 1978; 272: 549-50.
    CrossrefPubMedGoogle Scholar
  • 19 Fleury V, Loyau S, Lijnen HR. et al. Molecular assembly of plasminogen and tissue-type plasminogen activator on an evolving fibrin surface. Eur J Biochem 1993; 216: 549-56.
    CrossrefPubMedGoogle Scholar
  • 20 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.
    PubMedGoogle Scholar
  • 21 Nesheim M, Walker J, Wang W. et al. Modulation of fibrin cofactor activity in plasminogen activation. Ann N Y Acad Sci 2001; 936: 247-60.
    PubMedGoogle Scholar
  • 22 Weitz JI, Leslie B, Ginsberg J. Soluble fibrin degradation products potentiate tissue plasminogen activator-induced fibrinogen proteolysis. J Clin Invest 1991; 87: 1082-90.
    CrossrefPubMedGoogle Scholar
  • 23 Walker JB, Nesheim ME. A kinetic analysis of the tissue plasminogen activator and DSPAalpha1 cofactor activities of untreated and TAFIa-treated soluble fibrin degradation products of varying size. J Biol Chem 2001; 276: 3138-48.
    CrossrefPubMedGoogle Scholar
  • 24 Stewart RJ, Fredenburgh JC, Rischke JA. et al. Thrombin-activable fibrinolysis inhibitor attenuates (DD)E-mediated stimulation of plasminogen activation by reducing the affinity of (DD)E for tissue plasminogen activator A potential mechanism for enhancing the fibrin specificity of tissue plasminogen activator. J Biol Chem 2000; 275: 36612-20.
    CrossrefPubMedGoogle Scholar
  • 25 Bajzar L, Manuel R, Nesheim ME. Purification and characterization of TAFI, a thrombin-activable fibrinolysis inhibitor. J Biol Chem 1995; 270: 14477-84.
    CrossrefPubMedGoogle Scholar
  • 26 Bajzar L, Nesheim ME, Tracy PB. The profibrinolytic effect of activated protein C in clots formed from plasma is TAFI-dependent. Blood 1996; 88: 2093-100.
    PubMedGoogle Scholar
  • 27 Walker JB, Hughes B, James I. et al. Stabilization versus inhibition of TAFIa by competitive inhibitors in vitro. J Biol Chem 2003; 278: 8913-21.
    CrossrefPubMedGoogle Scholar
  • 28 Bajzar L, Kalafatis M, Simioni P. et al. An antifibrinolytic mechanism describing the prothrombotic effect associated with factor V Leiden. J Biol Chem 1996; 271: 22949-52.
    CrossrefPubMedGoogle Scholar
  • 29 Colucci M, D’Aprile AM, Italia A. et al. Thrombin activatable fibrinolysis inhibitor (TAFI) does not inhibit in vitro thrombolysis by pharmacological concentrations of t-PA. Thromb Haemost 2001; 85: 661-6.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Klement P, Liao P, Bajzar L. A novel approach to arterial thrombolysis. Blood 1999; 94: 2735-43.
    PubMedGoogle Scholar
  • 31 Nagashima M, Werner M, Wang M. et al. An inhibitor of activated thrombin-activatable fibrinolysis inhibitor potentiates tissue-type plasminogen activator-induced thrombolysis in a rabbit jugular vein thrombolysis model. Thromb Res 2000; 98: 333-42.
    CrossrefPubMedGoogle Scholar
  • 32 Mattsson C, Bjorkman JA, Abrahamsson T. et al. Local proCPU (TAFI) activation during thrombolytic treatment in a dog model of coronary artery thrombosis can be inhibited with a direct, small molecule thrombin inhibitor (melagatran). Thromb Haemost 2002; 87: 557-62.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 33 Sakharov DV, Barrertt-Bergshoeff M, Hekkenberg RT. et al. Fibrin-specificity of a plasminogen activator affects the efficiency of fibrinolysis and responsiveness to ultrasound: comparison of nine plasminogen activators in vitro. Thromb Haemost 1999; 81: 605-12.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 34 Colucci M, Cavallo LG, Agnelli G. et al. Properties of chimeric (tissue-type/urokinasetype) plasminogen activators obtained by fusion at the plasmin cleavage site. Thromb Haemost 1993; 69: 466-72.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 35 Kluft C, Nieuwenhuis HK, Rijken DC. et al. alpha 2-Antiplasmin Enschede: dysfunctional alpha 2-antiplasmin molecule associated with an autosomal recessive hemorrhagic disorder. J Clin Invest 1987; 80: 1391-400.
    CrossrefPubMedGoogle Scholar
  • 36 Rijken DC, Groeneveld E, Kluft C. et al. Alpha 2-antiplasmin Enschede is not an inhibitor, but a substrate, of plasmin. Biochem J 1988; 255: 609-15.
    PubMedGoogle Scholar
  • 37 Billing SClason, Meijer P, Kluft C. et al. Specific determination of plasmin inhibitor activity in plasma: documentation of specificity of manual and automated procedures. Blood Coagul Fibrinolysis 1999; 10: 487-94.
    CrossrefPubMedGoogle Scholar
  • 38 Schneider M, Nesheim M. Reversible inhibitors of TAFIa can both promote and inhibit fibrinolysis. J Thromb Haemost 2003; 01: 147-154.
    CrossrefPubMedGoogle Scholar
  • 39 Brouwers GJ, Vos HL, Leebeek FW. et al. A novel, possibly functional, single nucleotide polymorphism in the coding region of the thrombin-activatable fibrinolysis inhibitor (TAFI) gene is also associated with TAFI levels. Blood 2001; 98: 1992-3.
    CrossrefPubMedGoogle Scholar
  • 40 Gils A, Alessi MC, Brouwers E. et al. Development of a genotype 325-specific proCPU/TAFI ELISA. Arterioscler Thromb Vasc Biol 2003; 23: 1122-7.
    CrossrefPubMedGoogle Scholar
  • 41 Hortin GL, Gibson BL, Fok KF. Alpha 2-antiplasmin’s carboxy-terminal lysine residue is a major site of interaction with plasmin. Biochem Biophys Res Commun 1988; 155: 591-6.
    CrossrefPubMedGoogle Scholar
  • 42 Wang H, Yu A, Wiman B, Pap S. Identification of amino acids in antiplasmin involved in its noncovalent ‘lysine-binding-site’-dependent interaction with plasmin. Eur J Biochem 2003; 270: 2023-9.
    CrossrefPubMedGoogle Scholar