Partial Purification of a Specific Plasminogen Activator from Porcine Parotid Glands

Summary

A method is described for the extraction with buffers of near physiological pH of a plasminogen activator from porcine salivary glands. Substantial purification of the activator was achieved although this was to some extent complicated by concomitant extraction of nucleic acid from the glands. Preliminary characterization experiments using specific inhibitors suggested that the activator functioned by a similar mechanism to that proposed for urokinase, but with some important kinetic differences in two-stage assay systems. The lack of reactivity of the pig gland enzyme in these systems might be related to the tendency to protein-protein interactions observed with this material.

^* Present address : Searle Research Laboratories, High Wycombe, Bucks, England.

• References

• 1 Alkjaersig N, Fletcher A. P, Sherry S. E- aminocaproic acid - an inhibitor of plasminogen activation. Journal of Biological Chemistry 1959; 234: 832
• 2 Alkjaersig N, Fletcher A. P, Sherry S. The activation of human plasminogen; II. A kinetic study of activation with trypsin, urokinase and streptokinase. Journal of Biological Chemistry 1958; 233: 86
• 3 Ali S. Y, Lack C. M. Distribution of activator and proteolytic activity in the subcellular fractions of rabbit kidney. Biochemical Journal 1965; 96: 63
• 4 Andrews P. Estimation of molecular weights of proteins by Sephadex gel-filtration. Biochemical Journal 1965; 91: 222
• 5 Astrup T, Sterndorff I. The plasminogen activator in animal tissue. Acta physiologica Scandinavica 1956; 36: 250
• 6 Buckell M. The effect of citrate on euglobulin methods of estimating fibrinolytic activity. Journal of Clinical Pathology 1958; 17: 282
• 7 Cartwright T. The plasminogen activator of vampire bat saliva. Blood 1974; 43: 317
• 8 Hawkey C. M. Plasminogen activator in the saliva of the vampire bat. Nature 1966; 211: 434
• 9 KoK P, Astrup T. Isolation and purification of a tissue plasminogen activator and its comparison with urokinase. Biochemistry 1969; 8: 79
• 10 Lowry O. H, Rosebrough M. J, Farr A. C, Randall A. P. Protein measurement with the folin phenol reagent. Journal of Biological Chemistry 1951; 193: 265
• 11 Lanchantin G. F, Friedmann J. A, Hart D. W. Instruction of Soybean Trypsin Inhibition with Thrombin and its effect on Prothrombin Activation. Journal of Biological Chemistry 1969; 244: 865
• 12 Pugatch E. M, Foster E. A, MacFarlane D. E, Poole J. C. F. The extraction and separation of activators and inhibitors of fibrinolysis from bovine endothelium and mesothelium. British Journal of Haematology 1970; 18: 669
• 13 Rickli E. E, Bounok I, Zaugg H, Nitschmann H. S. Solublization of a tissuebound activator of plasminogen under physiological conditions. Chimica 1967; 21: 462
• 14 Rickli E. E, Zaugg H. Isolation and purification of highly enriched tissue activator of plasminogen from pig heart. Thrombosis et Diathesis Haemorrhagica 1970; 23: 64
• 15 Pobbins K. C, Summaria L, Elwyn D, Barlow G. M. Further studies on the purification and characterization of human plasminogen and plasmin. Journal of Biological Chemistry 1965; 240: 541
• 16 Spittle C. P, Lansdown N. M, Hawkey C. M. Micro modification of the dilute blood clot lysis time for determining fibrinolytic activity. Journal of Clinical Pathology 1968; 21: 780
• 17 Walton P. L. An improved fibrin plate method for assay of plasminogen activators. Clínica Chemica Acta 1966; 13: 680
• 18 Warburg O, Christian W. Isolierung und Kristallisation des Gärungsferments Enolase. Biochemische Zeitschrift 1942; 310: 384