Tridegin, a Novel Peptidic Inhibitor of Factor XIIIa from the Leech, Haementeria ghilianii, Enhances Fibrinolysis In Vitro

Summary

Tridegin is a potent inhibitor of factor Xllla from the leech, Haementeria ghilianii, which inhibits protein cross-linking. It modifies plasmin-mediated fibrin degradation as shown by the absence of D-dimer and approximately halves the time for fibrinolysis. Plasma clots formed in the presence of Tridegin lyse more rapidly when either streptokinase, tissue plasminogen activator or hementin is added 2 h after clot formation. The effect of Tridegin is markedly increased if clots are formed from platelet-rich plasma. Platelet-rich plasma clots are lysed much more slowly by the fibrinolytic enzymes used and if Tridegin is present, the rate of lysis returns almost to that of platelet- free clots. These studies indicate the important role of platelets in conferring resistance to commonly used fibrinolytic enzymes and suggest that protein cross-linking is an important step in this effect. Moreover they indicate that Tridegin, a small polypeptide, may have potential as an adjunct to thrombolytic therapy.

• References

• 1 McDonagh J. Structure and function of factor XIII. In: Haemostasis and Thrombosis: Basic Principles and Clinical Practice. Third edition Colman RW, Hirsh J, Marder VJ, Salzman EW. eds. J B Lippincott Company; Philadelphia, PA: 1994. pp 301-313
• 2 Schwartz ML, Pizzo SV, Hill RL, McKee PA. The effect of fibrin-stabilizing factor on the subunit structure of human fibrin. J Clin Invest 1971; 50: 1506-1513
• 3 Sakata Y, Aoki N. Significance of cross-linking of α₂-plasmin inhibitor to fibrin in inhibition of fibrinolysis and in hemostasis. J Clin Invest 1982; 69: 536-542
• 4 Jansen JWCM, Haverkate F, Koopman J, Nieuwenhuis HK, Kluft C, Boschman ThAC. Influence of factor Xllla activity on human whole blood clot lysis in vitro. Thromb Haemost 1987; 57: 171-175
• 5 van Giezen JJJ, Minkema J, Bouma N, Jansen JWCM. Cross-linking of α₂-antiplasmin to fibrin is a key factor in regulating blood clot lysis: species differences. Blood Coag Fibrinolysis 1993; 4: 869-875
• 6 Francis CW, Marder VJ. Rapid formation of large molecular weight α-polymers in cross-linked fibrin induced by high factor XIII concentrations: role of platelet factor XIII. J Clin Invest 1987; 80: 1459-1465
• 7 Fukue H, Anderson K, McPhedran P, Clyne L, McDonagh J. A unique factor XIII inhibitor to a fibrin-binding site on factor XIIIA. Blood 1992; 79: 65-74
• 8 Finney S, Seale L, Sawyer RT, Wallis RB. Tridegin, a new peptidic inhibitor of factor Xllla, from the blood sucking leech, Haementeria ghilianii . Biochem J. (in press).
• 9 Sawyer RT, Powell-Jones C, Munro R. The biological function of Hementin in the proboscis of the leech, Haementeria ghilianii . Blood Coag Fibrinolysis 1991; 2: 153-159
• 10 Tymiak AA, Tutle JG, Kimball SD, Wang T, Lee VG. A simple and rapid screen for inhibitors of factor Xllla. J Antibiotics 1993; 46: 204-206
• 11 Slaughter TF, Achyuthan KE, Lai T-S, Greenberg CS. A microtitre plate transglutaminase assay utilizing 5’-(biotinamido)pentylamine as substrate. Anal Biochem 1992; 205: 166-171
• 12 Swadesh JK, Huang IY, Budzynski AZ. Hementin, a fibrinogenolytic protease from the leech Haementeria ghilianii . J Chromatog 1990; 502: 359-369
• 13 Jones AJS, Meunier AM. A precise and rapid microtitre plate clot lysis assay: methodology, kinetic modelling and measurement of catalytic constants for plasminogen activation during fibrinolysis. Thromb Haemost 1990; 64: 455-463
• 14 Hauptmann J, Glusa E. Differential effects of staphylokinase, streptokinase and tissue-type plasminogen activator on the lysis of retracted human plasma clots and fibrinolytic plasma parameters in vitro. Blood Coag Fibrinolysis 1995; 6: 579-583
• 15 McKee PA, Mattock P, Hill RL. Subunit structure of human fibrinogen, soluble fibrin and cross-linked insoluble fibrin. Proc Natl Acad Sci USA 1970; 66: 738-744
• 16 Reed GL, Matsueda GR, Haber E. Platelet factor XIII increases the fibrinolytic resistance of platelet-rich clots by accelerating the crosslinking of α₂-antiplasmin. Thromb Haemost 1992; 68: 315-320
• 17 Purves L, Purves M, Brandt W. Cleavage of fibrin-derived D-dimer into monomers by endopeptidase from puff adder venom (Bitis arietans) acting at cross-linked sites of the γ-chain sequence of carboxy terminal cyanogen bromide γ-chain fragments. Biochemistry 1987; 26: 4640-466
• 18 Gabriel DA, Muga K, Boothroyd EM. The effect of fibrin structure on fibrinolysis. J Biol Chem 1992; 267: 24259-24263
• 19 Blomback B, Carlsson K, Hessel B, Liljeborg A, Procyk R, Aslund N. Native fibrin gel networks observed by 3D microscopy, permeation and turbidity. Biochim Biophys Acta 1989; 997: 096-110
• 20 Carr ME, Alving BM. Effect of fibrin structure on plasmin-mediated dissolution of plasma clots. Blood Coag Fibrinolysis 1995; 6: 567-573
• 21 Budzynski A. Interaction of hementin with fibrinogen and fibrin. Blood Coag Fibrinolysis 1991; 2: 149-152
• 22 Malinconico SM, Katz JB, Budzynski AZ. Fibrinogen degradation by hementin, a fibrinogenolytic anticoagulant from the salivary glands of the leech Haementeria ghilianii . J Lab Clin Med 1984; 104: 842-854
• 23 Sprengers ED, Kluft C. Plasminogen activator inhibitors. Blood 1987; 69: 381-387
• 24 Munro R, Powell-Jones C, Sawyer RT. Effects of hementin on fibrinogen mediated platelet aggregation/deaggregation in vitro. Platelets 1991; 2: 55-56
• 25 Shebuski RJ, Sitko GR, Claremon DA, Baldwin JJ, Remy DC, Stem AM. Inhibition of factor Xllla in a canine model of coronary thrombosis: effect on reperfusion and acute reocclusion after recombinant tissue-type plasminogen activator. Blood 1990; 75: 1455-1459
• 26 Leidy EM, Stem AM, Friedman PA, Bush LR. Enhanced thrombolysis by a factor Xllla inhibitor in a rabbit model of femoral artery thrombosis. Thromb Res 1990; 59: 15-26