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Thromb Haemost 1995; 73(05): 829-834
DOI: 10.1055/s-0038-1653876
Original Articles
Fibrinolysis
Schattauer GmbH Stuttgart

Localization of a Vitronectin Binding Region of Plasminogen Activator Inhibitor-1

Jaya Padmanabhan
2   The Department of Internal Medicine, The Bowman Gray School of Medicine, Winston-Salem, NC, USA
,
David C Sane
2   The Department of Internal Medicine, The Bowman Gray School of Medicine, Winston-Salem, NC, USA
› Author Affiliations
Further Information

Publication History

Received 23 August 1994

Accepted after resubmission 23 January 1995

Publication Date:
26 July 2018 (online)

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Summary

The PAI-1 binding site for VN was studied using two independent methods. PAI-1 was cleaved by Staph V8 protease, producing 8 fragments, only 2 of which bound to [125I]-VN. These fragments were predicted to overlap between residues 91-130. Since PAI-2 has structural homology to PAI-1, but does not bind to vitronectin, chimeras of PAI-1 and PAI-2 were constructed. Four chimeras, containing PAI-1 residues 1-70,1-105,1-114, and 1-167 were constructed and expressed in vitro. PAI-1, PAI-2, and all of the chimeras retained inhibitory activity for t-PA, but only the chimera containing PAI-1 residues 1-167 formed a complex with VN. Together, these results predict that the VN binding site of PAI-1 is between residues 115-130.

 

  • References

  • 1 Danφ K, Andreasen PA, Grondahl-Hansen J, Kristensen P, Nielsen LS, Skriver L. Plasminogen activators, tissue degradation and cancer. Adv Cancer Res 1985; 44: 139-266
    PubMedGoogle Scholar
  • 2 Loskutoff DJ, Sawdey M, Mimuro J. Type 1 plasminogen activator inhibitor. Prog Haemostas Thromb 1989; 9: 87-115
    PubMedGoogle Scholar
  • 3 Hekman CM, Loskutoff DJ. Bovine plasminogen activator inhibitor 1: specificity determinations and comparisons of the active, latent and guanidine- activated forms. Biochemistry 1988; 27: 2911-2918
    CrossrefPubMedGoogle Scholar
  • 4 Levin EG, Santell L. Conversion of active to latent plasminogen activator inhibitor from human endothelial cells. Blood 1987; 70: 1090-1098
    PubMedGoogle Scholar
  • 5 Kooistra T, Sprengers ED, van Hinsbergh VW M. Rapid inactivation of the plasminogen activator inhibitor upon secretion from cultured human endothelial cells. Biochem J 1986; 239: 497-503
    CrossrefPubMedGoogle Scholar
  • 6 Declerck PJ, de MolM, Alessi M-C, Baudner S, Pâques E-P, Preissner KT, Miiller-Berghaus G, Collen D. Purification and characterization of a plasminogen activator inhibitor 1 binding protein from human plasma. J Biol Chem 1988; 263: 15454-15461
    PubMedGoogle Scholar
  • 7 Wiman B, Almqvist Å, Sigurdardottir O, Lindahl T. Plasminogen activator inhibitor (PAI) is bound to vitronectin in plasma. FEBS Lett 1988; 242: 125-128
    CrossrefPubMedGoogle Scholar
  • 8 Salonen EM, Vaheri A, Pollanen J, Stephens R, Andreasen P, Mayer M, Dano K, Gailet J, Ruoslahti E. Interaction of plasminogen activator inhibitor (PAI-1) with vitronectin. J Biol Chem 1989; 264: 6339-6343
    PubMedGoogle Scholar
  • 9 Mimuro J, Loskutoff DJ. Purification of a protein from bovine plasma that binds to type 1 plasminogen activator inhibitor and prevents its interaction with the extracellular matrix. J Biol Chem 1989; 264: 936-939
    PubMedGoogle Scholar
  • 10 Seiffert D, Loskutoff D. Evidence that type 1 plasminogen activator inhibitor binds to the somatomedin B domain of vitronectin. J Biol Chem 1991; 266: 2824-2830
    PubMedGoogle Scholar
  • 11 Mimuro J, Muramatsu S, Kurano Y, Uchida Y, Ikadai H, Watanabe S, Sakata Y. Identification of the plasminogen activator inhibitor-1 binding heptapeptide in vitronectin. Biochemistry 1993; 32: 2314-2320
    CrossrefPubMedGoogle Scholar
  • 12 Kost C, Stüber W, Ehrlich HJ, Pannekoek H, Preissner KT. Mapping of binding sites for heparin, plasminogen activator inhibitor-1, and plasminogen to vitronectin’s heparin-binding region reveals a novel vitronectin- dependent feedback mechanism for the control of plasmin formation. J Biol Chem 1992; 267: 12098-12105
    PubMedGoogle Scholar
  • 13 Gechtman Z, Sharma R, Kreizman T, Fridkin M, Shaltiel S. Synthetic peptides derived from the sequence around the plasmin cleavage site in vitronectin. Use in mapping the PAI-1 binding site FEBS Lett 1993; 315: 293-297
    CrossrefPubMedGoogle Scholar
  • 14 Sisk WP, Davis GL, Kingsley D, Chiu AT, Reilly TM. High level synthesis of biologically active human plasminogen activator inhibitor 1 (PAI-1) in Escherichia coli. Gene 1990; 96: 305-309
    CrossrefPubMedGoogle Scholar
  • 15 Ye RD, Wun T-C, Sadler JE. cDNA cloning and expression in Escherichia coli of a plasminogen activator inhibitor from human placenta. J Biol Chem 1987; 262: 3718-3725
    PubMedGoogle Scholar
  • 16 Matsudaira P. Sequence from picomole quantities of protein electroblotted onto polyvinylidene difluoride membranes. J Biol Chem 1987; 262: 10035-10038
    PubMedGoogle Scholar
  • 17 Yatohgo T, Izumi M, Kashiwagi H, Hayashi M. Novel purification of vitronectin from human plasma by heparin affinity chromatography. Cell Struct Funct 1988; 13: 281-292
    CrossrefPubMedGoogle Scholar
  • 18 Reilly TM, Seetharam R, Duke JL, Davis GL, Pierce SK, Walton HL, Kingsley D, Sisk WP. Purification and characterization of recombinant plasminogen activator inhibitor-1 from Escherichia coli. J Biol Chem 1990; 265: 9570-9574
    PubMedGoogle Scholar
  • 19 Andreasen PA, Georg B, Lund LR, Riccio A, Stacey SN. Plasminogen activator inhibitors: hormonally regulated serpins. Mol Cell Endocrinol 1990; 68: 1-19
    CrossrefPubMedGoogle Scholar
  • 20 Huber R, Carrell RW. Implications of the three dimensional structure of α1-antitrypsin for structure and function of serpins. Biochemistry 1989; 28: 8951-8966
    CrossrefPubMedGoogle Scholar
  • 21 Fay WP, Owen WG. Platelet plasminogen activator inhibitor: purification and characterization of interaction with plasminogen activators and activated protein C. Biochemistry 1989; 28: 5773-5778
    CrossrefPubMedGoogle Scholar
  • 22 Declerck PJ, De Mol M, Vaughan DE, Collen D. Identification of a conformationally distinct form of plasminogen activator inhibitor-1, acting as a non-inhibitory substrate for tissue-type plasminogen activator. J Biol Chem 1992; 267: 11693-11696
    PubMedGoogle Scholar
  • 23 Radtke K-P, Wenz K-H, Heimburger N. Isolation of plasminogen activator inhibitor-2 (PAI-2) from human placenta. Evidence for vitronectin/PAI-2 complexes in human placenta extract. Biol Chem Hoppe-Seyler 1990; 371: 1117-1119
    PubMedGoogle Scholar
  • 24 Lawrence DA, Berkenpas MB, Palaniappan S, Ginsburg D. Localization of vitronectin binding domain in plasminogen activator inhibitor-1. J Biol Chem 1994; 269: 15223-15228
    PubMedGoogle Scholar
  • 25 Keijer J, Linders M, van ZonneveldA, Ehrlich HJ, de BoerJ, Pannekoek H. The interaction of plasminogen activator inhibitor 1 with plasminogen activators (tissue-type and urokinase type) and fibrin: localization of interaction sites and physiologic relevance. Blood 1991; 78: 401-409
    CrossrefPubMedGoogle Scholar
  • 26 Wagner OF, de VriesC, Hohmann C, Veerman H, Pannekoek H. Interaction between plasminogen activator inhibitor type 1 (PAI-1) bound to fibrin and either tissue-type activator (t-PA) or urokinase-type plasminogen activator (u-PA). Binding of t-PA/PAI-1 complexes to fibrin mediated by both the finger and the kringle-2 domain of t-PA J Clin Invest 1989; 84: 647-645
    CrossrefPubMedGoogle Scholar
  • 27 Ehrlich HJ, Gebbink RK, Keijer J, Linders M, Preissner KT, Pannekoek H. Alteration of serpin specificity by a protein cofactor. Vitronectin endows plasminogen activator inhibitor 1 with thrombin inhibitory properties J Biol Chem 1990; 265: 13029-13035
    PubMedGoogle Scholar