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Thromb Haemost 1999; 82(03): 1127-1131
DOI: 10.1055/s-0037-1614341
Letters to the Editor
Schattauer GmbH

Modulation of Cell-associated Plasminogen Activation by Stromelysin-1 (MMP-3)

F. Ugwu
1   From the Center for Molecular and Vascular Biology, University of Leuven, Belgium
,
G. Lemmens
1   From the Center for Molecular and Vascular Biology, University of Leuven, Belgium
,
D. Collen
1   From the Center for Molecular and Vascular Biology, University of Leuven, Belgium
,
H. R. Lijnen
1   From the Center for Molecular and Vascular Biology, University of Leuven, Belgium
› Author Affiliations
Further Information

Publication History

Received 13 April 1999

Accepted after revision 20 May 1999

Publication Date:
09 December 2017 (online)

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Summary

Stromelysin-1 (MMP-3) cleaves a 55 kDa kringle 1-4 fragment, containing the lysine-binding site(s) involved in cellular binding, from 92 kDa plasminogen and removes a 17 kDa NH2-terminal fragment, containing the cellular receptor-binding site, from 45 kDa urokinase (u-PA), but a potential role of MMP-3 in the regulation of cellular fibrinolytic activity by affecting binding and/or activation of plasminogen and/or single-chain u-PA has not been established. Human plasminogen (input concentration 100 nM for 4 × 106 cells per ml) was shown to bind specifically to human monocytoid THP-1 cells, to murine MMP-3 deficient smooth muscle cells (SMC) and fibroblasts (1.9, 0.92 and 1.0 × 106 molecules per cell, respectively). Treatment with MMP-3 (final concentration 0-50 nM) of cells saturated with bound plasminogen (about 25 nM), overnight at 37° C, resulted in a dose-dependent reduction of the amount of u-PA activatible plasminogen (reduction to 25-40% of the value in the absence of MMP-3). Immunoblotting with specific monoclonal antibodies and autoradiography of eluates of the cells treated with MMP-3 revealed cleavage of plasminogen into the 55 kDa fragment and miniplasminogen (kringle 5 plus the proteinase domain).

Binding of human single chain u-PA (scu-PA) to human THP-1 and HT 1080 cells amounted to 2.5 × 106and 7.1 × 106 molecules per cell, respectively. Treatment with MMP-3 (final concentration 0-25 nM) of cell-bound u-PA (about 17 nM for THP-1 and 47 nM for HT1080 cells), overnight at 37° C, did not alter cell-associated u-PA activity, measured in a direct chromogenic substrate assay or in a plasminogen-coupled chromogenic substrate assay (residual u-PA activity always ≥85% of that without MMP-3 treatment). Autoradiography of 125I-labeled u-PA moieties, removed from the cells by treatment with acid or with phosphatidylinositol phospholipase C, confirmed that u-PA remained essentially intact after MMP-3 treatment.

These data indicate that MMP-3 may downregulate cell-associated plasmin activity by decreasing the amount of activatible plasminogen, without affecting cell-bound u-PA activity.

 

  • References

  • 1 Lijnen HR, Collen D. Mechanisms of physiological fibrinolysis. Baillière’s Clin Haematol 1995; 8: 277-90.
    PubMedGoogle Scholar
  • 2 Collen D, Lijnen HR. Basic and clinical aspects of fibrinolysis and thrombolysis. Blood 1991; 78: 3114-24.
    PubMedGoogle Scholar
  • 3 Blasi F. Urokinase and urokinase receptor: a paracrine/autocrine system regulating cell migration and invasiveness. BioEssays 1993; 15: 105-11.
    CrossrefPubMedGoogle Scholar
  • 4 Plow EF, Félez J, Miles LA. Cellular regulation of fibrinolysis. Thromb Haemost 1991; 66: 32-6.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Miles LA, Dahlberg CM, Levin EG, Plow EF. Gangliosides interact directly with plasminogen and urokinase and may mediate binding of these components to cells. Biochemistry 1989; 28: 9337-43.
    CrossrefPubMedGoogle Scholar
  • 6 Miles LA, Dahlberg CM, Plescia J, Félez J, Kato K, Plow EF. Role of cell-surface lysines in plasminogen binding to cells: identification of alpha-enolase as a candidate plasminogen receptor. Biochemistry 1991; 30: 1682-91.
    CrossrefPubMedGoogle Scholar
  • 7 Hajjar KA, Jacovina AT, Chacko J. An endothelial cell receptor for plasminogen/tissue plasminogen activator. I. Identity with annexin II. J Biol Chem 1994; 269: 21191-7.
    PubMedGoogle Scholar
  • 8 Félez J, Miles LA, Fábregas P, Jardi M, Plow EF, Lijnen HR. Characterization of cellular binding sites and interactive regions within reactants required for enhancement of plasminogen activation by t-PA on the surface of leukocytic cells. Thromb Haemost 1996; 76: 577-84.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 9 Appella E, Robinson EA, Ullrich SJ, Stoppelli MP, Corti A, Cassani G, Blasi F. The receptor-binding sequence of urokinase. A biological function for growth factor module of proteases. J Biol Chem 1987; 262: 4437-40.
    PubMedGoogle Scholar
  • 10 Magdolen V, Rettenberger P, Koppitz M, Goretzki L, Kessler H, Weidle UH, Konig B, Graeff H, Schmitt M, Wilhelm O. Systematic mutational analysis of the receptor-binding region of the human urokinase-type plasminogen activator. Eur J Biochem 1996; 237: 743-51.
    CrossrefPubMedGoogle Scholar
  • 11 Ellis V, Behrendt N, Danø K. Plasminogen activation by receptor-bound urokinase. A kinetic study with both cell-associated and isolated receptor. J Biol Chem 1991; 266: 12752-8.
    PubMedGoogle Scholar
  • 12 Ellis V, Scully MF, Kakkar VV. Plasminogen activation initiated by single-chain urokinase-type plasminogen activator. Potentiation by U937 monocytes. J Biol Chem 1989; 264: 2185-8.
    PubMedGoogle Scholar
  • 13 Woessner Jr JF. Matrix metalloproteinases and their inhibitors in connective tissue remodelling. FASEB J 1991; 5: 2145-54.
    CrossrefPubMedGoogle Scholar
  • 14 Birkedal-Hansen H. Proteolytic remodeling of extracellular matrix. Curr Opinion Cell Biol 1995; 7: 728-35.
    CrossrefPubMedGoogle Scholar
  • 15 Matrisian LM. Metalloproteinases and their inhibitors in matrix remodeling. Trends Genet 1990; 6: 121-5.
    CrossrefPubMedGoogle Scholar
  • 16 Keski-Oja J, Lohi J, Tuuttila A, Tryggvason K, Vartio T. Proteolytic processing of the 72,000-Da Type IV collagenase by urokinase plasminogen activator. Exp Cell Res 1992; 202: 471-6.
    CrossrefPubMedGoogle Scholar
  • 17 Marcotte PA, Kozan IM, Dorwin SA, Ryan JM. The matrix metalloproteinase pump-1 catalyses formation of low molecular weight (pro)urokinase in cultures of normal human kidney cells. J Biol Chem 1992; 267: 13803-6.
    PubMedGoogle Scholar
  • 18 Dong Z, Kumar R, Yang X, Fidler IJ. Macrophage-derived metalloelastase is responsible for the generation of angiostatin in Lewis lung carcinoma. Cell 1997; 88: 801-10.
    CrossrefPubMedGoogle Scholar
  • 19 Patterson BC, Sang QA. Angiostatin-converting enzyme activities of human matrilysin (MMP-7) and gelatinase B/type IV collagenase (MMP-9). J Biol Chem 1997; 272: 28823-5.
    CrossrefPubMedGoogle Scholar
  • 20 Lijnen HR, Ugwu F, Bini A, Collen D. Generation of an angiostatin-like fragment from plasminogen by stromelysin-1 (MMP-3). Biochemistry 1998; 37: 4699-702.
    CrossrefPubMedGoogle Scholar
  • 21 Ugwu F, Van Hoef B, Bini A, Collen D, Lijnen HR. Proteolytic cleavage of urokinase-type plasminogen activator by stromelysin-1 (MMP-3). Biochemistry 1998; 37: 7231-6.
    CrossrefPubMedGoogle Scholar
  • 22 Lijnen HR, Van Hoef B, Collen D. On the role of the carbohydrate side chains of human plasminogen in its interaction with α2-antiplasmin and fibrin. Eur J Biochem 1981; 120: 149-54.
    CrossrefPubMedGoogle Scholar
  • 23 Lijnen HR, Van Hoef B, Collen D. Comparative kinetic analysis of the activation of human plasminogen by natural and recombinant single-chain urokinase-type plasminogen activator. Biochim Biophys Acta 1986; 884: 402-8.
    CrossrefPubMedGoogle Scholar
  • 24 Suzuki K, Enghild JJ, Morodomi T, Salvesen G, Nagase H. Mechanisms of activation of tissue procollagenase by matrix metalloproteinase 3 (stromelysin). Biochemistry 1990; 29: 10261-70.
    CrossrefPubMedGoogle Scholar
  • 25 Lijnen HR, Van Hoef B, Beelen V, Collen D. Characterization of the murine plasma fibrinolytic system. Eur J Biochem 1994; 224: 863-71.
    CrossrefPubMedGoogle Scholar
  • 26 Lijnen HR, Van Hoef B, Collen D. Characterization of the murine plasminogen/urokinase-type plasminogen activator system. Eur J Biochem 1996; 241: 840-8.
    CrossrefPubMedGoogle Scholar
  • 27 Lijnen HR, Lasters I, Verstreken M, Collen D, Jespers L. Screening panels of monoclonal antibodies using phage-displayed antigen. Anal Biochem 1997; 248: 211-5.
    CrossrefPubMedGoogle Scholar
  • 28 Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76: 4350-4.
    CrossrefPubMedGoogle Scholar
  • 29 Fraker PJ, Speck Jr JC. Protein and cell membrane iodinations with a sparingly soluble chloramide, 1, 3, 4, 6-tetrachloro-3a, 6a-diphenylglycoluril. Biochem Biophys Res Commun 1978; 80: 849-57.
    CrossrefPubMedGoogle Scholar
  • 30 Lijnen HR, Silence J, Van Hoef B, Collen D. Stromelysin-1 (MMP-3)-independent gelatinase expression and activation in mice. Blood 1998; 91: 2045-53.
    PubMedGoogle Scholar
  • 31 Carmeliet P, Collen D. Development and disease in proteinase-deficient mice: role of the plasminogen, matrix metalloproteinase and coagulation system. Thromb Res 1998; 91: 255-85.
    CrossrefPubMedGoogle Scholar
  • 32 Miles LA, Dahlberg CM, Plow EF. The cell-binding domains of plasminogen and their function in plasma. J Biol Chem 1988; 263: 11928-34.
    PubMedGoogle Scholar
  • 33 Christensen U. The AH-site of plasminogen and two C-terminal fragments. Biochem J 1984; 223: 413-21.
    CrossrefPubMedGoogle Scholar