Inhibition of Urokinase by Complex Formation with Human Antithrombin III in Absence and Presence of Heparin

Inge Clemmensen

doi:10.1055/s-0038-1646735

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1978; 39(03): 616-563
DOI: 10.1055/s-0038-1646735

Original Article

Schattauer GmbH Stuttgart

Inhibition of Urokinase by Complex Formation with Human Antithrombin III in Absence and Presence of Heparin

Inge Clemmensen

The Department of Clinical Chemistry, Hvidovre Hospital, DK-2650 Hvidovre, Denmark

› Institutsangaben

Abstract

als PDF herunterladen

Summary

Human antithrombin III was purified from fresh human plasma by affinity chromatography on heparin-Sepharose^®, affinity chromatography on concanavalin A Sepharose^®, gel filtration on Ultrogel^® AcA 34, ion exchange chromatography on DEAE A-50 Sephadex^® and preparative agarose gel electrophoresis. The hydrolytic activity of urokinase (plasminogen activator from urine) on acetyl-glycyl-L-lysine methyl ester acetate (Ac-gly-lys-OMe Ac) was inhibited by antithrombin III in a slow time-dependent manner. Heparin accelerated the reaction between activator and inhibitor. Inhibition of catalytic activity was associated with the formation of an 1:1 molar complex between activator and inhibitor as revealed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The complex was also demonstrated by crossed Immunoelectrophoresis against anti-antithrombin III.

PDF (857 kb)

Referenzen

References
1 Abildgaard U, Fagerhol MK, Egeberg O. 1970; Comparison of progressive antithrombin activity and the concentrations of three thrombin inhibitors in human plasma. Scandinavian Journal of Clinical and Laboratory Investigation 26: 349
2 Aoki N, Von Kaulla KN. 1971; Human serum plasminogen antiactivator: its distinction from antiplasmin. American Journal of Physiology 220: 1137
3 Aoki N, Von Kaulla KN. 1971; The extraction of vascular plasminogen ativator from human cadavers and a description of some of its properties. American Journal of Clinical Pathology 55: 171
4 Astrup T. 1966; Tissue activators of plasminogen. Federation Proceedings 25: 42
5 Bachmann F, Fletcher AP, Alkjaersig N, Sherry S. 1964; Partial purification and properties of the plasminogen activator from pig heart. Biochemistry 3: 1578
6 Bernik MB, Kwan HC. 1967; Origin of fibrinolytic activity in cultures of the human kidney. Journal of Laboratory and Clinical Medicine 70: 650
7 Biggs R, Denson KWE, Akman N, Borrett R, Hadden M. 1970; Antithrombin 3, antifactor X_a and heparin. British Journal of Haematology 19: 283
8 Burrowes CE, Habal FM, Movat HZ. 1975; The inhibition of human plasma kallikrein by antithrombin III. Thrombosis Research 7: 175
9 Clemmensen I, Christensen F. 1976; Inhibition of urokinase by complex formation with human α₁-antitrypsin. Biochimica et biophysica acta 429: 591
10 Clemmensen L, Thorsen S, Müllertz S. 1976; Purification of a plasminogen activator inhibitor indistinguishable from α₁-antitrypsin and an urokinase inhibitor in pregnancy plasma. Haemostasis 5: 218
11 Collen D, Cock FD, Verstraete M. 1975; Immunochemical distinction between antiplasmin and alpha₁-antitrypsin. Thrombosis Research 7: 245
12 Collen D. 1976; Identification and some properties of a new fast-reacting plasmin inhibitor in human plasma. European Biochimistry 69: 209
13 Crawford GPM, Ogston D. 1975; The action of antithrombin III on plasmin and activators of plasminogen. Biochimica et biophysica acta 391: 189
14 Gallimore MJ. 1975; Serum inhibitors of fibrinolysis. British Journal of Haematology 31: 217
15 Ganrot PO. 1972; Crossed Immunoelectrophoresis. Scandinavian Journal of Clinical and Laboratory Investigations 29 Suppl (Suppl. 124) 39
16 Hedner U. 1973; Studies on an inhibitor of plasminogen activation in human serum. Thrombosis et Diathesis Haemorrhagica 30: 414
17 Heimburger N. 1975. Proteinase inhibitors of human plasma-their properties and control function. In: Reich E, Rifkin DB, Shaw E. (Eds.) Proteases and Biological Control. Cold Spring Harbor Laboratory; USA: 367
18 Highsmith RF, Rosenberg RD. 1974; The inhibition of human plasmin by human antithrombin-heparin cofactor. Journal of Biological Chemistry 249: 4335
19 Kawano T, Morimoto K, Uemura Y. 1970; Partial purification and properties of urokinase inhibitor from human placenta. The Journal of Biochemistry 67: 333
20 Lahiri B, Bagdasarien A, Mitchell B, Talamo RC, Colman RW, Rosenberg RD. 1976; Antithrombin-Heparin Cofactor: An Inhibitor of Plasma Kallikrein. Archives of Biochemistry and Biophysics 175: 737
21 Landmann H, Markwardt F. 1970; Irreversible synthetische Inhibitoren der Urokinase. Experientia 26: 145
22 Laurell CB. 1972; Electroimmuno assay. Scandinavian Journal of Clinical and Laboratory Investigations 29 Suppl (Suppl. 124) 39
23 Moroi M, Aoki N. 1976; Isolation and characterization of α₂-plasmin inhibitor from human plasma. Journal of Biological Chemistry 251: 5956
24 Müllertz S. 1972; Molecular forms of plasmin and protease inhibitors in human fibrinolytic postmortem plasma. Scandinavian Journal of Clinical and Laboratory Investigations 20: 113
25 Müllertz S. 1974; Different molecular forms of plasminogen and plasmin produced by urokinase in human plasma and their relation to protease inhibitors and lysis of fibrinogen and fibrin. Biochemical Journal 143: 273
26 MüLLERTZ S, Clemmensen I. 1976; The primary inhibitor of plasmin in human plasma. Biochemical Journal 159: 545
27 Ong EB, Johnson AL, Schoellmann P. 1976; Identification of an active site histidine in urokinase. Biochimica et biophysica acta 429: 252
28 Rimon A, Shamash Y, Shapiro B. 1966; The Plasmin inhibitor of human plasma. Journal of Biological Chemistry 241: 5102
29 Robbins KC, Bernabe P, Arzadan L, Summaria L. 1973; The histidine loop of human plasmin: light (B) chain active center histidine sequence. Journal of Biological Chemistry 248: 1631
30 Rosenberg RD. 1974; The effect of heparin on factor XI_a and plasmin. Thrombosis et Diathesis Haemorrhagica 33: 51
31 Rosenberg RD, Damus PS. 1973; The purification and mechanism of action of human antithrombin-heparin cofactor. Journal of Biological Chemistry 248: 6490
32 Sawjer WD, Fletcher AP, Alkjaersig N. 1960; Studies on the thrombolytic activity of human plasma. Journal of Clinical Investigation 39: 426
33 Seegers WH, Cole ER, Harmison CR, Monkhouse FC. 1964; Neutralization of autoprothrombin C activity by antithrombin. Canadian Journal of Biochemistry 42: 359
34 Stead N, Kaplan AP, Rosenberg RD. 1976; Inhibition of activated factor XII by antithrombin-heparin cofactor. Journal of Biological Chemistry 251: 6481
35 Takeda Y, Parkhill TR, Nakabayashi M. 1972; Effects of heparin and E-ammocaproide acid in dogs on plasmin-¹²⁵I generation in response to urokinase injections and venous injury. Journal of Clinical Investigation 51: 2678
36 Telesforo P, Semeraro M, Verstrate M, Collen C. 1975; Inhibition of plasmin by antithrombin III-heparin complex in vitro in human plasma and during streptokinase therapy in man. Thrombosis Research 7: 669
37 Uszynsky M, Abildgaard U. 1971; Separation and characterization of two fibrinolytic inhibitors from human placenta. Thrombosis et Diathesis Haemorrhagica 25: 580
38 Vennerød AM, Laake K. 1975; Inhibition of purified plasma kallikrein by antithrombin III and heparin. Thrombosis Research 7: 223
39 White WF, Barlow GH, Mozen MM. 1966; The isolation and characterization of plasminogen activators (urokinase) from human urine. Biochemistry 5: 2160
40 Yin ET, Wessler S, Stoll P. 1971; Identity of plasma-activated factor X inhibitor with antithrombin III and heparin cofactor. Journal of Biological Chemistry 246: 3712