Download PDF
Thromb Haemost 1990; 64(02): 270-275
DOI: 10.1055/s-0038-1647299
Original Article
Schattauer GmbH Stuttgart

Effect of Peptides on the Inactivation of Tissue Plasminogen Activator by Plasminogen Activator Inhibitor-1 and on the Binding of Tissue Plasminogen Activator to Endothelial Cells

A J Wittwer
The Department of Cell Culture and Bochemistry, Monsanto Co., St. Louis, MO, U.S.A
,
M A Sanzo
The Department of Cell Culture and Bochemistry, Monsanto Co., St. Louis, MO, U.S.A
› Author Affiliations
Further Information

Publication History

Received 24 November 1989

Accepted after revision07 May 1990

Publication Date:
25 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

The effectiveness of tissue plasminogen activator (tPA) in thrombolytic therapy is dependent upon the rate at which therapeutically administered tPA reaches the clot site and the proportion of that tPA which is enzymatically active. Interactions between tPA and its main plasma inhibitor (PAI-1) and between tPA and the endothelial cells lining blood vessels are two factors which may limit efficacy. In an attempt to identify the regions of the tPA molecule involved in these interactions, we have examined a series of synthetic peptides with amino acid sequences corresponding to different regions of the tPA molecule for their ability to protect tPA from inactivation by PAI-1 and for their ability to reduce the binding of tPA to endothelial cells. Three peptides were identified which were especially effective at maintaining tPA activity in the presence of PAI-1 and three others were found which had a lesser effect. These same peptides were also found to inhibit the binding of tPA to endothelial cells. This suggests that the same regions of the tPA molecule are involved in both processes. None of the peptides inhibited the binding of tPA to fibrin. These peptides may serve as models for the development of agents for enhancing the activity of both endogenous tPA and of tPA administered in thrombolytic therapy.

 

  • References

  • 1 Nilsson I, Ljungnér H, Tengborn L. Two different mechanisms in patients with venous thrombosis and defective fibrinolysis: low concentrations of plasminogen activator or increased concentration of plasminogen activator inhibitor. Br Med J 1985; 290: 1453-1456
    CrossrefPubMedSearch in Google Scholar
  • 2 Jørgensen M, Bonnevie-Nielsen V. Increased concentration of the fact-acting plasminogen activator inhibitor in plasma associated with familial venous thrombosis. Br J Haematol 1987; 65: 175-180
    CrossrefPubMedSearch in Google Scholar
  • 3 Juhan-Vague I, Valadier J, Alessi MC, Aillaud MF, Ansaldi J, Philip-Joet C, Holvoet P, Serradimigni A, Collen D. Deficient tPA release and elevated PA inhibitor levels in patients with spontaneous or recurrent deep venous thrombosis. Thromb Haemostas 1987; 57: 67-72
    PubMedSearch in Google Scholar
  • 4 Mehta J, Mehta P, Lawson P, Saldeen T. Plasma tissue plasminogen activator inhibitor levels in coronary artery disease: correlation with age and serum triglyceride concentrations. J Am Cell Cardiol 1987; 9: 263-268
    CrossrefPubMedSearch in Google Scholar
  • 5 Hamsten A, de Faire U, Walldius G, Dahlen G, Szamosi A, Landou C, Blombäck M, Wiman B. Plasminogen activator inhibitor in plasma: a risk factor for recurrent myocardial infarction. Lancet 1987; 2: 3-9
    PubMedSearch in Google Scholar
  • 6 Kruithof EK O, Nicolosa G, Bachmann F. Plasminogen activator inhibitor 1: development of a radioimmunoassay and observations on its plasma concentration during venous occlusion and after platelet aggregation. Blood 1987; 70: 1645-1653
    PubMedSearch in Google Scholar
  • 7 Booth NA, Anderson JA, Bennett B. Platelet release protein which inhibits plasminogen activators. J Clin Pathol 1985; 38: 825-830
    CrossrefPubMedSearch in Google Scholar
  • 8 Sprengers ED, Akkerman JW, Jansen BG. Blood platelet plasminogen activator inhibitor: two different pools of endothelial cell type plasminogen activator inhibitor in human blood. Thromb Haemostas 1986; 55: 325-329
    PubMedSearch in Google Scholar
  • 9 Jang I-K, Gold HK, Ziskind AA, Fallon JT, Holt RE, Leinbach RC, May JW, Collen D. Differential sensitivity of erythrocyte-rich and platelet-rich arterial thrombi to lysis with recombinant tissue-type plasminogen activator. Circulation 1989; 79: 920-928
    CrossrefPubMedSearch in Google Scholar
  • 10 Hajjar KA, Hamel NM, Harpel PC, Nachman RL. Binding of tissue plasminogen activator to cultured human endothelial cells. J Clin Invest 1987; 80: 1712-1719
    CrossrefPubMedSearch in Google Scholar
  • 11 Bamathan E, Kuo A, Van der Keyl H, McCrae KR, Larsen G, Clines DB. Tissue-type plasminogen activator binding to human endothelial cells. J Biol Chem 1988; 263: 7792-7797
    PubMedSearch in Google Scholar
  • 12 Beebe DP. Binding of tissue plasminogen activator to human umbilical vein endothelial cells. Thromb Res 1987; 46: 241-254
    CrossrefPubMedSearch in Google Scholar
  • 13 Sakata Y, Okada M, Noro A, Matsuda M. Interaction of tissue-type plasminogen activator and plasminogen activator inhibitor 1 on the surface of endothelial cells. J Biol Chem 1988; 263: 1960-1969
    PubMedSearch in Google Scholar
  • 14 Ranby M, Wallen P. A sensitive parabolic rate assay for the tissue plasminogen activator. Prog Fibrinolysis 1981; 5: 233-235
    PubMedSearch in Google Scholar
  • 15 Ranby M. Studies on the kinetics of plasminogen activation by tissue plasminogen activator. Biochim Biophys Acta 1982; 704: 461-469
    CrossrefPubMedSearch in Google Scholar
  • 16 Katagiri K, Okada K, Hattori H, Yano M. Bovine endothelial cell plasminogen activator inhibitor. Purification and heat activation. Eur J Biochem 1988; 176: 81-87
    CrossrefPubMedSearch in Google Scholar
  • 17 Rijken P, Hoylaerts M, Collen D. Fibrinolytic properties of one-chain and two-chain extrinsic (tissue-type) plasminogen activator. J Biol Chem 1982; 257: 2920-2925
    PubMedSearch in Google Scholar
  • 18 Hekman CM, Loskutoff DJ. Bovine plasminogen activator inhibitor 1: specificity determinations and comparison of the active, latent, and guanine-activated forms. Biochemistry 1988; 27: 2911-2918
    CrossrefPubMedSearch in Google Scholar
  • 19 Ny T, Elgh F, Lund B. The structure of the human tissue-type plasminogen activator gene: Correlation of intron and exon structures to functional and structural domains. Proc Natl Acad Sci USA 1984; 81: 5355-5359
    CrossrefPubMedSearch in Google Scholar
  • 20 Morton PA, Owensby DA, Sobel BE, Schwartz AL. Catabolism of tissue-type plasminogen activator by the human hepatoma cell line Hep G2. Modulation by plasminogen activator inhibitor type I. J Biol Chem 1989; 264: 7228-7235
    PubMedSearch in Google Scholar
  • 21 Mimuro J, Schleef RR, Loskutoff DJ. Extracellular matrix of cultured bovine aortic endothelial cells contains functionally active type 1 plasminogen activator inhibitor. Blood 1987; 70: 721-728
    PubMedSearch in Google Scholar
  • 22 Mimuro J, Loskutoff DJ. Binding of type 1 plasminogen activator inhibitor to the extracellular matrix of cultured bovine endothelial cells. J Biol Chem 1989; 264: 5058-5063
    PubMedSearch in Google Scholar
  • 23 Declerck PJ, De Mol M, Alessi M-C, Baudner S, Paques E-P, Preissner KT, Mliller-Berghaus G, Collen D. Purification and characterization of a plasminogen activator inhibitor 1 binding protein from human plasma. Identification as a multimeric form of S protein (vitronectin). J Biol Chem 1988; 263: 15454-15461
    PubMedSearch in Google Scholar
  • 24 Salonen E-M, Vaher A, Pollanen J, Stephens R, Andreasen P, Mayer M, Danø K, Gailit J, Ruoslahti E. Interaction of plasminogen activator inhibitor (PAI-1) with vitronectin. J Biol Chem 1989; 264: 6339-6343
    PubMedSearch in Google Scholar
  • 25 Mimuro J, Loskutoff DJ. Purification of a protein a from bovine plasma that binds to type 1 plasminogen activator inhibitor and prevents its interaction with extracellular matrix. Evidence that the protein is vitronectin. J Biol Chem 1989; 264: 936-939
    PubMedSearch in Google Scholar
  • 26 Wun T-C, Palmier MO, Siegel NR, Smith CE. Affinity purification of active plasminogen activator inhibitor-1 (PAI-1) using immobilized anhydrourokinase. Demonstration of the binding, stabilization, and activation of PAI-1 by vitronectin. J Biol Chem 1989; 264: 7862-7868
    PubMedSearch in Google Scholar
  • 27 Chmielewska J, Ranby M, Wiman B. Kinetics of the inhibition of plasminogen activators by the plasminogen-activator inhibitor. Evidence of “second-site” interactions. Biochem J 1988; 251: 327-332
    CrossrefPubMedSearch in Google Scholar
  • 28 Masson C, Angles-Cano E. Kinetic analysis of the interaction between plasminogen activator inhibitor-1 and tissue-type plasminogen activator. Biochem J 1988; 256: 237-244
    CrossrefPubMedSearch in Google Scholar
  • 29 Verheijen JH, Caspers MP M, Chang GT G, de Munk GA W, Pouwels PH, Enger-Valk BE. Involvement of finder domain and kringle 2 domain of tissue-type plasminogen activator in fibrin binding and stimulation of activity by fibrin. EMBO J 1986; 5: 3525-3530
    PubMedSearch in Google Scholar
  • 30 Appella E, Robinson EA, Ullrich SJ, Stoppelli MP, Corti A, Casani G, Blasi F. The receptor-binding sequence of urokinase. A biological function for the growth-factor module of proteases. J Biol Chem 1987; 262: 4437-4440
    PubMedSearch in Google Scholar