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Thromb Haemost 1989; 62(03): 909-916
DOI: 10.1055/s-0038-1651027
Original Article
Schattauer GmbH Stuttgart

The Nature of Synergy between Tissue-Type and Single Chain Urokinase-Type Plasminogen Activators

Edward T A Fry
The Cardiovascular Division, Washington University School of Medicine, St. Louis, Missouri, USA
,
David L Mack
The Cardiovascular Division, Washington University School of Medicine, St. Louis, Missouri, USA
,
Burton E Sobel
The Cardiovascular Division, Washington University School of Medicine, St. Louis, Missouri, USA
› Author Affiliations
Further Information

Publication History

Received: 04 October 1988

Accepted after revision 24 May 1989

Publication Date:
30 June 2018 (online)

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Summary

Enhancement of thrombolysis with combinations of tissue-type and single chain urokinase plasminogen activators (t-PA and scu-PA) has been demonstrated in vivo but has not been seen consistently in vitro. This study was designed to characterize interactions between t-PA and scu-PA with respect to rate of and extent of thrombolysis in vitro and to delineate mechanisms responsible. Combinations of t-PA and scu-PA at selected concentrations synergistically enhanced thrombolysis in vitro compared with thrombolysis induced by either activator alone. Enhanced thrombolysis did not occur at the expense of fibrin specificity since the extent of fibrinogenolysis and consumption of α2-antiplasmin were significantly less with synergistic combinations of t-PA and scu-PA compared with equi-effective concentrations of either activator alone. Attenuation of complex formation of t-PA and two chain u-PA (tcu-PA), formed from scu-PA, with plasma proteins did not appear to contribute to enhancement of thrombolysis as assessed by fibrin autography. Binding of 125I-t-PA to thrombi was increased by 27% at 1 hr and by 21% at 2 hr in the presence of scu-PA (p <0,001 for both). Conversion of scu-PA to tcu-PA was enhanced when thrombi were exposed to scu-PA in the presence of t-PA. Results of this study indicate that t-PA and scu-PA at selected concentrations enhance thrombolysis in vitro synergistically without compromising fibrin specificity. Enhanced binding of t-PA to thrombi in the presence of scu-PA and enhanced conversion of scu-PA to tcu-PA appear to contribute to synergy between t-PA and scu-PA for thrombolysis.

Keywords

Tissue-type plasminogen activator (t-PA) - Single chain urokinase plasminogen activator (scu-PA) - Synergism - Thrombolysis
 

  • References

  • 1 Sobel BE. Coronary thrombosis with tissue-type plasminogen activator (t-PA): Emerging strategies. J Am Coll Cardiol 1986; 8: 1220-1225
    CrossrefPubMedGoogle Scholar
  • 2 Verstraete M, Collen D. Thrombolytic therapy for the eighties. Blood 1986; 67: 1529-1541
    PubMedGoogle Scholar
  • 3 Collen D, Bounameaux H, DeCock F, Lijnen HR, Verstraete M. Analysis of coagulation and fibrinolysis during intravenous infusion of recombinant human tissue-type plasminogen activator in patients with acute myocardial infarction. Circulation 1986; 73: 511-517
    CrossrefPubMedGoogle Scholar
  • 4 Van de Werf F, Vanhaecke J, DeGeest H, Verstraete M, Collen D. Coronary thrombolysis with recombinant single-chain urokinase-type plasminogen activator in patients with acute myocardial infarction. Circulation 1986; 74: 1066-1070
    CrossrefPubMedGoogle Scholar
  • 5 Collen D, Stassen JM, Stump DC, Verstraete M. Synergism of thrombolytic agents in vivo. Circulation 1986; 74: 838-842
    CrossrefPubMedGoogle Scholar
  • 6 Collen D, Van de Werf F. Coronary arterial thrombolysis with low- dose synergistic combinations of recombinant tissue-type plasminogen activator (rt-PA) and recombinant single-chain urokinase-type plasminogen activator (rscu-PA) for acute myocardial infarction. Am J Cardiol 1987; 60: 431-434
    CrossrefPubMedGoogle Scholar
  • 7 Collen D, DeCock F, Demarsin E, Lijnen HR, Stump DC. Absence of synergism between tissue-type plasminogen activator (t-PA), single-chain urokinase-type plasminogen activator (scu-PA) and urokinase on clot lysis in a plasma milieu in vitro. Thromb Haemostas 1986; 56: 35-39
    PubMedGoogle Scholar
  • 8 Gurewich V, Panned R. A comparative study of the efficacy and specificity of tissue plasminogen activator and pro-urokinase: Demonstration of synergism and of different thresholds of non-selectivity. Thromb Res 1986; 44: 217-228
    CrossrefPubMedGoogle Scholar
  • 9 Pannell R, Black J, Gurewich V. Complementary modes of action of tissue-type plasminogen activator and pro-urokinase by which their synergistic effect on clot lysis may be explained. J Clin Invest 1988; 81: 853-859
    CrossrefPubMedGoogle Scholar
  • 10 Fry ET A, Sobel BE. Lack of interference by heparin with thrombolysis or binding of tissue-type plasminogen activator to thrombi. Blood 1988; 71: 1347-1352
    PubMedGoogle Scholar
  • 11 Rampling MW, Gaffney PJ. The sulfite precipitation method for fibrinogen measurements: Its use on small samples in the presence of fibrinogen degradation products. Clin Chem Acta 1976; 67: 43-52
    CrossrefPubMedGoogle Scholar
  • 12 Gaffney PJ, Brasher M, Lord K, Kirkwood TB L. Activation of plasminogen as a feature in its assay. Haemostasis 1977; 6: 72-88
    PubMedGoogle Scholar
  • 13 Lucore CL, Sobel BE. Interactions of tissue-type plasminogen activator with plasma inhibitors and their pharmacologic implications. Circulation 1988; 77: 660-669
    CrossrefPubMedGoogle Scholar
  • 14 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
    CrossrefPubMedGoogle Scholar
  • 15 Hunter WM, Greenwood FC. Preparation of 131I-labeled human growth hormone of high specific activity. Nature 1962; 194: 495-496
    CrossrefPubMedGoogle Scholar
  • 16 Jespersen J, Astrup T. A study of fibrin plate assay of fibrinolytic agents. Haemostasis 1983; 13: 301-315
    PubMedGoogle Scholar
  • 17 Berenbaum MC. The expected effect of a combination of agents: The general solution. J Theor Biol 1985; 114: 413-431
    CrossrefPubMedGoogle Scholar
  • 18 United Nations Scientific Committee on the Effects of Atomic Radiation. Ionizing Radiation: Sources and Biological Effects. Report to the General Assembly. UNSCEAR 1982; Publ. E.82.IX 8: 727-765
    PubMedGoogle Scholar
  • 19 Chou JC, Talalay P. Analysis of combined drug effects: A new look at a very old problem. Trends in Pharmacol 1983; 4: 450-454
    PubMedGoogle Scholar
  • 20 Chou JC, Talalay P. Quantitative analysis of dose-effect relationships: The combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 1984; 22: 27-55
    CrossrefPubMedGoogle Scholar
  • 21 Stump DC, Thienpont M, Collen D. Urokinase-related proteins in human urine. J Biol Chem 1986; 261: 1267-1273
    PubMedGoogle Scholar
  • 22 Stump DC, Lijnen HR, Collen D. Biochemical and biological properties of single-chain urokinase-type plasminogen activator. Cold Spring Harbor Symp Quant Biol 1986; 51: 563-569
    CrossrefPubMedGoogle Scholar
  • 23 Ranby M, Wallen P. Enzymatic properties of tissue-type plasminogen activator. In: Thrombolysis: Biological and Therapeutic Properties of new Thrombolytic Agents. Collen D, Lijnen HR, Verstraete M. (eds). Churchill Livingstone; Edinburgh: 1985
    Google Scholar
  • 24 Hoylaerts M, Rijken DC, Lijnen HR, Collen D. Kinetics of the activation of plasminogen by human tissue plasminogen activator. J Biol Chem 1982; 257: 2912-2919
    PubMedGoogle Scholar
  • 25 Gurewich V, Pannell R. Synergism of tissue-type plasminogen activator (t-PA) and single-chain urokinase-type plasminogen activator (scu-PA) on clot lysis in vitro and a mechanism for this effect. Thromb Haemostas 1987; 57: 372
    PubMedGoogle Scholar
  • 26 Collen D. Rebuttal. Synergism of tissue-type plasminogen activator (t-PA) and single-chain urokinase-type plasminogen activator (scu-PA) on clot lysis in vitro and a mechanism of this effect. Thromb Haemostas 1987; 57: 373
    PubMedGoogle Scholar
  • 27 Strieker RB, Wong D, Tak Shiu D, Reyes PT, Shuman MA. Activation of plasminogen by tissue plasminogen activator on normal and thrombasthenic platelets: Effects on surface proteins and platelet aggregation. Blood 1986; 68: 275-280
    PubMedGoogle Scholar
  • 28 Blasi F. Surface receptors for urokinase plasminogen activator. Fibrinolysis 1988; 2: 73-84
    PubMedGoogle Scholar
  • 29 Gurewich V. Experiences with pro-urokinase and potentiation of its fibrinolytic effect by urokinase and by tissue plasminogen activator. J Am Coll Cardiol 1987; 10: 16B-21B
    CrossrefPubMedGoogle Scholar
  • 30 Topol EJ, Califf RM, George BS, Kereiakes DJ, Rothbaum D, Candela RJ, Abbotsmith CW, Pinkerton CA, Stump DC, Collen D, Lee KL, Pitt B, Kline EM, Boswick JM, O'Neill WW, Stack RS. the TAMI Study Group Coronary arterial thrombolysis with combined infusion of recombinant tissue-type plasminogen activator and urokinase in patients with acute myocardial infarction. Circulation 1988; 77: 1100-1107
    CrossrefPubMedGoogle Scholar
  • 31 van Zonneveld AJ, Veerman H, Pannekoek J. Autonomous functions of structural domains on human tissue-type plasminogen activator. Proc Natl Acad Sci USA 1986; 83: 4670-4674
    CrossrefPubMedGoogle Scholar
  • 32 Lijnen HR, Zamarron C, Blabert M, Winkler ME, Collen D. Activation of plasminogen by pro-urokinase. J Bio Chem 1986; 261: 1253-1258
    PubMedGoogle Scholar
  • 33 Petersen LC, Lund LR, Nielsen LS, Dano K, Skriver L. One-chain urokinase-type plasminogen activator from human sarcoma cells is a proenzyme with little or no intrinsic activity. J Biol Chem 1988; 263: 11189-11195
    PubMedGoogle Scholar
  • 34 Ellis V, Scully MF, Kakkar VV. Plasminogen activation by singlechain urokinase in functional isolation. J Biol Chem 1987; 262: 14998-15003
    PubMedGoogle Scholar
  • 35 Lijnen HR, Gheysen D, de Foresta F, Pierard L, Jacobs P, Bollen A, Collen D. Characterisation of a mutant of recombinant human single chain urokinase-type plasminogen activator (scu-PA), obtained by substitution of arginine 156 and lysine 158 with threonine. Fibrinolysis 1988; 2: 85-93
    PubMedGoogle Scholar
  • 36 Gurewich V, Pannell R, Broeze RJ, Mao J. Characterization of the intrinsic fibrinolytic properties of pro-urokinase through a study of plasmin-resistant mutant forms produced by site-directed mutagenesis of lysine-158. J Clin Invest 1988; 82: 1956-1962
    CrossrefPubMedGoogle Scholar
  • 37 Rijken DC, Hoylaerts M, Collen D. Fibrinolytic properties of one-chain and two-chain human extrinsic (tissue-type) plasminogen activator. J Biol Chem 1982; 257: 2920-2925
    PubMedGoogle Scholar
  • 38 Collen D, Stump DC, Van de Werf F. Coronary thrombolysis in patients with acute myocardial infarction by intravenous infusion of synergic thrombolytic agents. Am Heart J 1986; 112: 38-44
    PubMedGoogle Scholar