Purification and Properties of an Antiactivator Fraction from Bovine Serum

Summary

A method is described for the purification of antiactivator from bovine euglobulin-free serum by means of gelfiltration and ion exchange chromatography. The purified antiactivator has no antifibrinolytic activity. It has a molecular weight of about 115,000 and it appears to be a gamma globulin. The dissociation constant of its complex with urokinase is 1.6 X 10^-9 M and the maximum urokinase binding is close to 2000 CTA units per mg. Its concentration in bovine serum is 0.37%. Flufenamate displaces urokinase from the antiactivator at very low concentrations, about 10^-4 M. Cysteine restores its activity if lost by standing. Also an antifibrinolysin fraction is obtained free of antiactivator activity.

• References

• 1 Aoki N, Von Kaulla KN. 1969; Inactivation of human serum plasminogen antiactivator by synthetic fibrinolysis inducers. Thrombosis et Diathesis Haemorrhagica 22: 251
• 2 Aoki N, Von Kaulla KN. 1971; Human serum plasminogen antiactivator: its distinction from antiplasmin. American Journal of Physiology 220: 1137
• 3 Aoki N, Moroi M. 1974; Distinction of serum inhibitor of activator-induced clot lysis from ax antitrypsin. Proceedings Society Experimental Biology and Medicine 146: 567
• 4 Bennett NB. 1967; A method for the quantitative assay of inhibitor of plasminogen activation in human serum. Thrombosis et Diathesis Haemorrhagica 17: 12
• 5 Berg W, Korsan-Bengtsen K, Ygge J. 1968; Theoretical bases and standardization of the one stage lysis time method for determination of urokinase. Thrombosis et Diathesis Haemorrhagica 19: 169
• 6 Cohn EJ, Edsall JT. 1943. Proteins, amino acids and peptides as ions and dipolar ions. Reinhold Publ. Comp; New York: p 428
• 7 Hedner U. 1973; Studies on an inhibitor of plasminogen activation in human serum. Thrombosis et Diathesis Haemorrhagica 30: 414
• 8 Hedner U. 1974; Fibrinolytic inhibitors. Thrombosis et Diathesis Haemorrhagica Suppl 59: 287
• 9 Laemmli UK. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680
• 10 Maxwell RE, Nickel VS, Lewandowski V. 1962; Preparations of plasminogen deficient fibrinogen and thrombin. Biochemical Biophysical Research Communications 7: 50
• 11 Nanninga LB, Guest MM. 1965; Blocking of inhibition of fibrinolysin by mercurials and oxidizing agents. Archives Biochemistry and Biophysics 109: 607
• 12 Nanninga LB, Guest MM. 1966; Rapid determination of antifibrinolysin (antiplasmin) in plasma. Thrombosis et Diathesis Haemorrhagica 15: 213
• 13 Nanninga LB, Guest MM. 1967; a Preparation and properties of anticoagulant split product of fibrinogen and its determination in plasma. Thrombosis et Diathesis Haemorrhagica 17: 440
• 14 Nanninga LB. 1967; b Rapid determination of profibrinolysin (plasminogen) in plasma. Thrombosis et Diathesis Haemorrhagica 17: 8
• 15 Nanninga LB, Guest MM. 1971; Antiactivator (antikinase) activity of human plasma and its blocking by parachloromercuribenzoate and salicylate. Thrombosis et Diathesis Haemorrhagica 26: 541
• 16 Nanninga LB. 1975; a Blocking of urokinase binding to plasma inhibitor by diiodosalicylate. Federation Proceedings 34: 290
• 17 Nanninga LB. 1975; b Molar concentrations of fibrinolytic components, especially free fibrinolysin in vivo. Thrombosis et Diathesis Haemorrhagica 33: 244
• 18 Nanninga LB. 1977; Soluble fibrin complexes in early fibrin digests. Thrombosis and Haemostasis 37: 192
• 19 Nanninga LB. 1979; The binding of diiodosalicylate and flufenamate to the plasma antiactivator: analysis of chemical fibrinolysis. Thrombosis and Haemostasis 41: 357
• 20 Neville DM, Glossman H. 1974; Molecular weight determination from discontinuous gels with sodium dodecylsulfate. Methods of Enzymology 32: 92
• 21 Schachman HK. 1957; Ultracentrifugation, diffusion and viscometry. Methods in Enzymology 4: 32