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

Ana H. C. Guimarães; Dingeman C. Rijken

doi:10.1160/TH03-06-0377

Thrombosis and Haemostasis, Inhaltsverzeichnis

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

¹Gaubius Laboratory, TNO Prevention and Health, Leiden

²the Department of Physiology, Institute for Cardiovascular Research, VU University, Amsterdam, The Netherlands

,

Dingeman C. Rijken

¹Gaubius Laboratory, TNO Prevention and Health, Leiden

³the Department of Hematology, Erasmus University Medical Center, Rotterdam,The Netherlands

› Institutsangaben

Abstract

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

Volltext

Referenzen

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.
2 Nesheim ME. TAFI. Fibrinolysis & Proteolysis 1999; 13: 72-77.
3 Bajzar L. Thrombin activatable fibrinolysis inhibitor and an antifibrinolytic pathway. Arterioscler Thromb Vasc Biol 2000; 20: 2511-8.
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.
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.
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.
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.
8 Marx PF, Dawson PE, Bouma BN. et al. Plasmin-mediated activation and inactivation of thrombin-activatable fibrinolysis inhibitor. Biochemistry 2002; 41: 6688-96.
9 Redlitz A, Tan AK, Eaton DL, Plow EF. Plasma carboxypeptidases as regulators of the plasminogen system. J Clin Invest 1995; 96: 2534-8.
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.
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.
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.
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.
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.
15 Christensen U. C-terminal lysine residues of fibrinogen fragments essential for binding to plasminogen. FEBS Lett 1985; 182: 43-46.
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.
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.
18 Wiman B, Collen D. Molecular mechanism of physiological fibrinolysis. Nature 1978; 272: 549-50.
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.
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.
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.
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.
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.
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.
25 Bajzar L, Manuel R, Nesheim ME. Purification and characterization of TAFI, a thrombin-activable fibrinolysis inhibitor. J Biol Chem 1995; 270: 14477-84.
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.
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.
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.
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.
30 Klement P, Liao P, Bajzar L. A novel approach to arterial thrombolysis. Blood 1999; 94: 2735-43.
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.
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.
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.
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.
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.
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.
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.
38 Schneider M, Nesheim M. Reversible inhibitors of TAFIa can both promote and inhibit fibrinolysis. J Thromb Haemost 2003; 01: 147-154.
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.
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.
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.
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.