The Antithrombotic Properties of Human Prothrombin Fragment 1.2 in Mice

Summary

We have investigated the antithrombotic properties of prothrombin fragment 1.2 (F_1.2) in this study. To do this, we established the minimum concentration of human placental tissue factor or human a-thrombin that was lethal in mice within 5 min after intravenous injection. Prothrombin F_1.2 protected the mice from the lethal effect of tissue factor or α-thrombin in a dose dependent manner, with 500 μg (14 nmoles) of prothrombin F_1.2 per mouse being the minimum amount required to protect all mice from the lethal effect of either thrombogenic stimulus. The minimum dose of heparin which protected mice from the lethal effect of thrombin or tissue factor was 6 units or ∼3.3 nmoles. The observation that prothrombin F_1.2 has antithrombotic properties suggests prothrombin F_1.2 can modulate coagulation in vivo, as it has previously been shown to do in vitro.

• References

• 1 Govers-Riemslag JW P, Speijer H, Zwaal RF A, Rosing J. The effects of bovine prothrombin fragment 1 and fragment 1.2 on prothrombin activation. Thromb Res 1985; 38: 375-88
• 2 Nawroth PP, Kisiel W, Stern DM. Anticoagulant and antithrombotic properties of a y-carboxyglutamic acid-rich peptide derived from the light chain of blood coagulation factor X. Thromb Res 1986; 44: 625-37
• 3 Wildgoose P, Kisiel W. Inhibition of prothrombin activation by factor X and factor IX Gla-peptides. Biochem Biophys Res Commun 1988; 152: 1207-12
• 4 Aronson DL, Stevan L, Ball AP, Franza Jr BR, Finlayson JS. Generation of the combined prothrombin activation peptide (F12) during the clotting of blood and plasma. J Clin Invest 1977; 60: 1410-8
• 5 Ofosu FA, Sie P, Modi GJ, Fernnndez F, Buchanan MR, Blajchman MA, Boneu B, Hirsh J. The inhibition of thrombin-dependent positive feedback-reactions is critical to the expression of the anticoagulant effect of heparin. Biochem J 1987; 243: 579-88
• 6 Bell WN, Alton HD. A brain extract as a substitute for platelet suspension in the thromboplastin generation test. Nature 1954; 174: 880
• 7 Beutler E, West C. The storage of hard-packed red blood cells in citrate-phosphate-dextrose (CPD) and CPD-Adenine (CPDA-1). Blood 1979; 54: 280-4
• 8 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5
• 9 Ofosu FA, Modi G, Cerskus AL, Hirsh J, Blajchman MA. Heparin with low affinity to antithrombin III inhibits the activation of prothrombin in normal plasma. Thromb Res 1982; 28: 487-97
• 10 Thomas L. Studies on the intravascular thromboplastic effect of tissue suspension in mice. 11. A factor in normal rabbit serum which inhibits the thromboplastic effect of the sedimentable tissue component. Bull John Hopkins Hospital 1947; 81: 26-42
• 11 Maim KG, Elion J, Butkowski RJ, Downing M, Nesheim ME. Prothrombin. In: Methods in Enzymology. Lorand L. (ed) Academic Press; New York: 1981. 80 286-302
• 12 Salzman EW, Hirsh J. Prevention of venous thromboembolism. In: Hemostasis and Thrombosis. Basic Principles and Clinical Practice Colman RW, Hirsh J, Marder VJ, Salzman EW. (eds) 2nd. edition J. B. Lippincott; Philadelphia, PA: 1987. p 1252
• 13 Wessler S. Medical management of venous thromboembolism. Ann Rev Med 1976; 27: 313-9
• 14 Wang L. Plasma volume, cell volume, total blood volume and F cells factor in the normal and splenectomized Sherman rat. Am J Physiol 1959; 196: 188-92
• 15 Teitel JM, Bauer KA, Lau HK, Rosenberg RD. Studies of the prothrombin activation pathway utilizing radioimmunoassays for the F2/F1 + 2 fragment and thrombin-antithrombin complex. J Biol Chem 1982; 59: 1086-97
• 16 Ofosu FA, Buchanan MR, Anvari N, Smith LM, Blajchman MA. Plasma anticoagulant mechanism of heparin, heparan sulfate and dermatan sulfate. Ann NY Acad Sci 1989; 556: 123-31
• 17 Nesheim ME, Abbott T, Jenny R, Mann KG. Evidence that the thrombin-catalyzed feedback cleavage of fragment 1.2 at Argl 54-Ser155 promotes the release of thrombin from the catalytic surface during the activation of bovine prothrombin. J Biol Chem 1988; 263: 1037-44