Thromb Haemost 1971; 25(01): 114-128
DOI: 10.1055/s-0038-1654285
Originalarbeiten – Original Articles – Travaux Originaux
Schattauer GmbH

Plasminogen-Fibrinogen Complex Formation as a Prelude to Fibrinogenolysis

A Density-Gradient Ultracentrifugation Study of Radioiodinated Systems Involving Urokinase, Plasminogen, and Fibrinogen*
Anne P. Ball
1   Departments of Surgery and of Microbiology-Immunology, Duke University Medical Center, Durham, N. C. 27706 (USA)
,
D Silver
1   Departments of Surgery and of Microbiology-Immunology, Duke University Medical Center, Durham, N. C. 27706 (USA)
,
Eugene D. Day
1   Departments of Surgery and of Microbiology-Immunology, Duke University Medical Center, Durham, N. C. 27706 (USA)
› Author Affiliations
Supported by Contract AT-(40-1)-3195 with the U.S. Atomic Energy Commission and by Grants HE-08929 and NB-06233 with the National Institutes of Health, USPHS
Further Information

Publication History

Publication Date:
28 June 2018 (online)

Summary

Reproducible density-gradient ultracentrifugation reference profiles of the ratezonal type were established individually for radioiodinated fibrinogen, plasminogen, and urokinase. Two- and three-component radioiodinated mixtures were then studied to determine whether stable complexes would form between any two components and thus change the density-gradient distribution profiles.

Urokinase did not form stable complexes with plasminogen, fibrinogen, or fibrin, nor with any plasminogen-fibrinogen or plasminogen-fibrin mixtures.

Plasminogen was found to form stable complexes with fibrinogen and with fibrin. Urokinase added to such mixtures resulted in fibrinogenolysis and fibrinolysis, but not in stable urokinase binding.

The data support the proposition that urokinase does not activate plasminogen directly but requires the formation of plasminogen-fibrin(ogen) complexes as a prelude (conformational change?) to plasmin conversion and subsequent fibrin(ogen)olysis.

 
  • References

  • 1 Alkjaersig N. The purification and properties of human plasminogen. Biochem. J 93: 171 1964;
  • 2 Alkjaersig N, Fletcher A. P, Sherry S. The activation of human plasminogen. I. Spontaneous activation in glycerol. J. biol. Chem 233: 81 1958;
  • 3 Alkjaersig N, Fletcher A. P, Sherry S. The activation of human plasminogen. II. Kinetic study of activation with trypsin, urokinase, and streptokinase. J. biol. Chem 233: 86 1958;
  • 4 Ambrus G, Back N, Ambrus J. L. Fibrinolytic effect of plasmin-anti—plasmin complexes. Fed. Proc 19: 58 1960;
  • 5 Ball A. P. Ph. D. Dissertation, Duke University; 1970
  • 6 Ball A. P, Day E. D. Urokinase-induced fibrinolysis of 125I-fibrin : Conditions for first-order activation kinetics, density-gradient separation of split products, and a microassay. Thrombos. Diathes. haemorrh. (Stuttg) 24: 463 1970;
  • 7 Ball A. P, Day E. D. Immunological identification of two urokinases. Thrombos. Diathes. haemorrh. (Stuttg) 24: 475 1970;
  • 8 Berg W. Plasminogen and urokinase determination based on the lysis time method. Acta med. scand. Suppl 478 1967;
  • 9 Berg W, Korsan-Bengtsen K, Ygge Y. Preparation of human plasminogen suitable as a substrate for quantitative determination of plasminogen activators. Thrombos. Diathes. haemorrh. (Stuttg) 15: 511 1966;
  • 10 Brakman P. Ph. D. Thesis, University of Amsterdam. 1967. published by Scheltema and Holkema N.V; Amsterdam:
  • 11 Budzynski A. Z, Stahl M, Kopec M, Latallo Z. S, Wegrzynowicz Z, Kowalski E. High molecular weight products of the late stage of fibrinogen proteolysis and their structure relation to the fibrin molecule. Biochim. biophys. Acta (Amst) 147: 313 1967;
  • 12 Davidson N, Wang J. G. Effects of density gradient centrifugation on equilibria involving macromolecules. In: Rich A, Davidson N. (eds.), Structural Chemistry and Molecular Biology. Freeman and Co; San Francisco: 1968
  • 13 Day E. D, Ball A. P. Immunological identification of three urokinases. Thromb. Diathes. haemorrh. (Stuttg). (In press). 1971
  • 14 Day E. D, Ball A. P, Jeffords D, Woodard Jr. W. T, Silver D. The immunoassay of human urokinase. Thromb. Diathes. haemorrh. (Stuttg) 21: 273 1969;
  • 15 Day E. D, Lassiter S, Fritz R. B. Kadioiodination of antibodies adsorbed to insoluble antigens. J. Immunol 98: 67 1967;
  • 16 Day E. D, Planinsek J, Korngold L, Pressman D. Tumor-localizing antibodies purified from antisera against Murphy rat lymphosarcoma. J. Nat. Cancer Inst 17: 517 1956;
  • 17 Day E. D, Planinsek J. A, Pressman D. Triadic labeling with 130I, 131I, and 133I for controlled determinations of tumor-localizing antibodies. J. Nat. Cancer Inst 26: 1321 1961;
  • 18 Doleschel W, Auerswald W. Determination of molecular weight of uroprotein fractions with urokinase activity by means of molecular sieving. Med. Pharmacol. Exper 16: 225 1967;
  • 19 Fritz R. B, Lassiter S, Day E. D. A quantitative precipitin study of porcine fibrinogen employing paired labels. J. Immunol 98: 1213 1967;
  • 20 Hamberg U. Studies on plasminogen activators. I. Sucrose density gradient distribution of activators from normal and lytic human plasmas. Acta chim. scand 20: 2539 1966;
  • 21 Hamberg U, Savolainen M-L. Studies on plasminogen activators. II. Method of separation of purified urokinase from small samples of urine. Acta chim. scand 20: 2551 1966;
  • 22 Iwanaga S, Wallen P, Gröhndahl M. J, Henschen A, Blombäck B. Isolation and characterization of N-terminal fragments obtained by plasmin digestion of human fibrinogen. Biochim. biophys. Acta (Amst) 147: 606 1967;
  • 23 Laki K. The polymerization of proteins: The action of thrombin on fibrinogen. Arch. Biochem 32: 317 1951;
  • 24 Lesuk A, Terminiello L, Traver J. Crystalline urokinase: Some properties. Science 147: 880 1965;
  • 25 Pressman D, Day E. D, Blau M. The use of paired labelling in the determination of tumorlocalizing antibodies. Cancer Res 17: 845 1957;
  • 26 Robbins K, Summaria L. Purification of human plasminogen and plasmin by gel filtration on Sephadex and chromatography on diethylaminoethyl-Sephadex. J. biol. Chem 238: 952 1963;
  • 27 Sherry S, Fletcher A. P, AIkjaersig N. Fibrinolysis and fibrinolytic activity in man. Physiol. Rev 39: 343 1959;
  • 28 Summaria L, Hsieh B, Robbins K. The specific mechanism of activation of human plasminogen to plasmin. J. biol. Chem 242: 4279 1967;
  • 29 Triantaphyllopoulos E. D. O. Fibrinogenolysis : The micromolecular derivatives. Brit. J. Haemat 15: 337 1968;
  • 30 White W. F, Barlow G. H, Mozen M. M. The isolation and characterization of plasminogen activators (urokinase) from human urine. Biochemistry 05: 2160 1966;
  • 31 Woodard Jr. W. T, Ball A. P, Day E. D, Silver D. Immunological assay of plasma from patients infused with urokinase. Surgical Forum XXI: 215 1970;
  • 32 Woodard W. T, Day E. D, Silver D. The fate of infused urokinase. Surgery 68: 692 1970;