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Thromb Haemost 1998; 79(06): 1171-1176
DOI: 10.1055/s-0037-1615036
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Schattauer GmbH

Regulation of Gelatinase Activity in Mice with Targeted Inactivation of Components of the Plasminogen/Plasmin System

H. R. Lijnen
From the Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
,
J. Silence
From the Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
,
G. Lemmens
From the Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
,
L. Frederix
From the Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
,
D. Collen
From the Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
› Author Affiliations
Further Information

Publication History

Received 16 December 1997

Accepted after revision 19 February 1998

Publication Date:
07 December 2017 (online)

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Summary

To investigate a potential physiological role of the plasminogen/ plasmin system in activation of the matrix metalloproteinase (MMP) system, the distribution of latent and active MMP-2 (gelatinase A) or MMP-9 (gelatinase B) was monitored in aorta extracts and in serum-free conditioned cell culture medium obtained from wild-type (WT) mice and from mice with deficiency of tissue-type plasminogen activator (t-PA–/–), urokinase-type plasminogen activator (u-PA–/–), plasminogen activator inhibitor-1 (PAI-1–/–) or plasminogen (Plg–/–).

In aorta extracts, the contribution of active MMP-2 to the total MMP-2 level ranged between 7 and 16% for the different genotypes, whereas active MMP-9 was not detected.

The contribution of active 58 kDa MMP-2 to the total MMP-2 level (active plus latent) ranged between 14 and 29% (mean of 3 experiments) for fibroblasts of the different genotypes, and between 18 and 32% for smooth muscle cells, and was relatively constant in time (7-72 h). The contribution of active 83 kDa MMP-9 to the total MMP-9 level ranged between 15 and 29% for fibroblasts of the different genotypes and was relatively constant in time (24-72 h); corresponding values were 17 to 57% for smooth muscle cells, with the exception of Plg–/– smooth muscle cells which had undetectable levels of active MMP-9. Addition of plasmin(ogen) to the cell culture medium of fibroblasts did not significantly affect the distribution of active and latent MMP-2, but resulted in an approximately two-fold enhancement of the contribution of active MMP-9. In macrophages of Plg–/– mice, active MMP-9 was detected only when the cells were cultured in the presence of plasminogen.

These data indicate that activation of proMMP-2 occurs independently of the physiological plasminogen activators and of plasmin(ogen) in all the cell types evaluated. Activation of proMMP-9 was enhanced in the presence of plasmin(ogen), but active MMP-9 was also detected in fibroblasts of Plg–/– mice, indicating that in vivo activation may occur via plasmin(ogen)-independent mechanisms.

 

  • References

  • 1 Matrisian LM. Metalloproteinases and their inhibitors in matrix remodeling. Trends Genet 1990; 6: 121-5.
    CrossrefPubMedGoogle Scholar
  • 2 Woessner Jr. JF. Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J 1991; 5: 2145-54.
    CrossrefPubMedGoogle Scholar
  • 3 Birkedal-Hansen H. Proteolytic remodeling of extracellular matrix. Curr Opinion in Cell Biol 1995; 7: 728-35.
    PubMedGoogle Scholar
  • 4 Murphy G. Matrix metalloproteinases and their inhibitors. Acta Orthop Scand Suppl 1995; 266: 55-60.
    PubMedGoogle Scholar
  • 5 Lijnen HR, Collen D. Mechanisms of physiological fibrinolysis. Baillière’s Clin Haematol 1995; 8: 277-90.
    PubMedGoogle Scholar
  • 6 Eeckhout Y, Vaes G. Further studies on the activation of procollagenase, the latent precursor of bone collagenase. Effects of lysosomal cathepsin B, plasmin and kallikrein, and spontaneous activation. Biochem J 1977; 166: 21-31.
    CrossrefPubMedGoogle Scholar
  • 7 He CS, Wilhelm SM, Pentland AP, Marmer BL, Grant GA, Eisen AZ, Goldberg GI. Tissue cooperation in a proteolytic cascade activating human interstitial collagenase. Proc Natl Acad Sci USA 1989; 86: 2632-6.
    CrossrefPubMedGoogle Scholar
  • 8 Okada Y, Gonoji Y, Naka K, Tomita K, Nakanishi I, Iwata K, Yamashita K, Hayakawa T. Matrix metalloproteinase 9 (92-kDa gelatinase/type IV collagenase) from HT 1080 human fibrosarcoma cells. Purification and activation of the precursor and enzymic properties. J Biol Chem 1992; 267: 21712-9.
    PubMedGoogle Scholar
  • 9 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
  • 10 Mazzieri R, Masiero L, Zanetta L, Monea S, Onisto M, Garbisa S, Mignatti P. Control of type IV collagenase activity by components of the urokinase-plasmin system: a regulatory mechanism with cell-bound reactants. EMBO J 1997; 16: 2319-32.
    CrossrefPubMedGoogle Scholar
  • 11 Keski-Oja J, Lohi J, Tuuttila A, Tryggvason K, Vartio T. Proteolytic processing of the 72,000-Da Type IV collagenase by urokinase plasminogen activation. Exp Cell Research 1992; 202: 471-6.
    CrossrefPubMedGoogle Scholar
  • 12 Ogata Y, Enghild JJ, Nagase H. Matrix metalloproteinase 3 (stromelysin) activates the precursor for the human matrix metalloproteinase 9. J Biol Chem 1992; 267: 3581-4.
    PubMedGoogle Scholar
  • 13 Imai K, Yokohama Y, Nakanishi I, Ohuchi E, Fujii Y, Nakai N, Okada Y. Matrix metalloproteinase 7 (matrilysin) from human rectal carcinoma cells. Activation of the precursor, interaction with other matrix metalloproteinases and enzymic properties. J Biol Chem 1995; 270: 6691-7.
    CrossrefPubMedGoogle Scholar
  • 14 Sato H, Takino T, Okada Y, Cao J, Shinagawa A, Yamamoto E, Seiki M. A matrix metalloproteinase expressed on the surface of invasive tumour cells. Nature 1994; 370: 61-5.
    CrossrefPubMedGoogle Scholar
  • 15 Knauper P, Smith B, Lopez Otin C, Murphy G. Activation of progelatinase B (proMMP-9) by active collagenase-3 (MMP-13). Eur J Biochem 1997; 248: 369-73.
    CrossrefPubMedGoogle Scholar
  • 16 Murphy G, Atkinson S, Ward R, Gavrilovic J, Reynolds JJ. The role of plasminogen activators in the regulation of connective tissue metalloproteinases. Ann NY Acad Sci 1992; 667: 1-12.
    CrossrefPubMedGoogle Scholar
  • 17 Carmeliet P, Schoonjans L, Kieckens L, Ream B, Degen J, Bronson R, De Vos R, van den Oord JJ, Collen D, Mulligan RC. Physiological consequences of loss of plasminogen activator gene function in mice. Nature 1994; 368: 419-24.
    CrossrefPubMedGoogle Scholar
  • 18 Carmeliet P, Kieckens L, Schoonjans L, Ream B, van Nuffelen A, Prendergast G, Cole M, Bronson R, Collen D, Mulligan RC. Plasminogen activator inhibitor-1 gene-deficient mice. I. Generation by homologous recombination and characterization. J Clin Invest 1993; 92: 2746-55.
    CrossrefPubMedGoogle Scholar
  • 19 Ploplis VA, Carmeliet P, Vazirzadeh S, Van Vlaenderen I, Moons L, Plow EF, Collen D. Effects of disruption of the plasminogen gene on thrombosis, growth, and health in mice. Circulation 1995; 92: 2585-93.
    CrossrefPubMedGoogle Scholar
  • 20 Giles AR. Guidelines for the use of animals in biomedical research. Thromb Haemost 1987; 58: 1078-84.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 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
  • 22 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
  • 23 Heussen C, Dowdle EB. Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Anal Biochem 1980; 102: 196-202.
    CrossrefPubMedGoogle Scholar
  • 24 Kleiner DE, Stetler Stevenson WG. Quantitative zymography: detection of picogram quantities of gelatinases. Anal Biochem 1994; 218: 325-9.
    CrossrefPubMedGoogle Scholar
  • 25 Dewerchin M, Van Nuffelen A, Wallays G, Bouché A, Moons L, Carmeliet P, Mulligan RC, Collen D. Generation and characterization of urokinase receptor-deficient mice. J Clin Invest 1996; 97: 870-8.
    CrossrefPubMedGoogle Scholar
  • 26 Lijnen HR, Moons L, Beelen V, Carmeliet P, Collen D. Biological effects of combined inactivation of plasminogen activator and plasminogen activator inhibitor-1 gene function in mice. Thromb Haemost 1995; 74: 1126-31.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 27 Carmeliet P, Moons L, Ploplis V, Plow E, Collen D. Impaired arterial neointima formation in mice with disruption of the plasminogen gene. J Clin Invest 1997; 99: 200-8.
    CrossrefPubMedGoogle Scholar
  • 28 Carmeliet P, Moons L, Herbert J-M, Crawley J, Lupu F, Lijnen HR, Collen D. Urokinase-type but not tissue-type plasminogen activator mediates arterial neointima formation in mice. Circ Res 1997; 81: 829-39.
    CrossrefPubMedGoogle Scholar
  • 29 Bendeck MP, Zempo N, Clowes AW, Galardy RE, Reidy MA. Smooth muscle cell migration and matrix metalloproteinase expression after arterial injury in the rat. Circ Res 1994; 75: 539-45.
    CrossrefPubMedGoogle Scholar
  • 30 Jackson CL, Reidy MA. The role of plasminogen activation in smooth muscle cell migration after arterial injury. Ann NY Acad Sci 1992; 667: 141-50.
    CrossrefPubMedGoogle Scholar
  • 31 Carmeliet P, Moons L, Lijnen HR, Baes M, Lemaître V, Tipping P, Drew A, Eeckhout Y, Shapiro S, Lupu F, Collen D. Urokinase-generated plasmin is a candidate activator of matrix metalloproteinases during atherosclerotic aneurysm formation. Nature Genet 1997; 17: 439-44.
    CrossrefPubMedGoogle Scholar
  • 32 Baramova EN, Bajou K, Remacle A, L’Hoir C, Krell HW, Weidle UH, Noel A, Foidart JM. Involvement of PA/plasmin system in the processing of pro-MMP-9 and in the second step of pro-MMP-2 activation. FEBS Lett 1997; 405: 157-62.
    CrossrefPubMedGoogle Scholar
  • 33 Murphy G, Ward R, Hembry RM, Reynolds JJ, Kuhn K, Tryggvason K. Characterization of gelatinase from pig polymorphonuclear leucocytes. A metalloproteinase resembling tumour type IV collagenase. Biochem J 1989; 258: 463-72.
    CrossrefPubMedGoogle Scholar
  • 34 Okada Y, Morodomi T, Enghild JJ, Suzuki K, Yasui A, Nakanishi I, Salvesen G, Nagase H. Matrix metalloproteinase 2 from human rheumatoid synovial fibroblasts. Purification and activation of the precursor and enzymic properties. Eur J Biochem 1990; 194: 721-30.
    CrossrefPubMedGoogle Scholar
  • 35 Chen JM, Aimes RT, Ward GR, Youngleib GL, Quigley JP. Isolation and characterization of a 70-kDa metalloprotease (gelatinase) that is elevated in Rous sarcoma virus-transformed chicken embryo fibroblasts. J Biol Chem 1991; 266: 5113-21.
    PubMedGoogle Scholar
  • 36 Moll UM, Youngleib GL, Rosinski KB, Quigley JP. Tumor promoter-stimulated Mr 92,000 gelatinase secreted by normal and malignant human cells: isolation and characterization of the enzyme from HT 1080 tumor cells. Cancer Res 1990; 50: 6162-70.
    PubMedGoogle Scholar
  • 37 Mignatti P, Robbins E, Rifkin DB. Tumor invasion through the human amniotic membrane: requirement for a proteinase cascade. Cell 1986; 47: 487-98.
    CrossrefPubMedGoogle Scholar
  • 38 Reich R, Thompson EW, Iwamoto Y, Martin GR, Deason JR, Fuller GC, Miskin R. Effects of inhibitors of plasminogen activator, serine proteinases, and collagenase IV on the invasion of basement membranes by metastatic cells. Cancer Res 1988; 48: 3307-12.
    PubMedGoogle Scholar
  • 39 Lijnen HR, Silence J, Van Hoef B, Collen D. Stromelysin-1 (MMP-3)-independent gelatinase expression and activation in mice. Blood. (in press).
    PubMedGoogle Scholar