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Myocardial matrix metalloproteinase(s): localization and activation

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Abstract

Matrix metalloproteinases (MMPs) and neutrophil elastate (NE) may each contribute to fibrillar collagen degradation in various disease states. Little, however, is known about the activation and localization of MMP in the heart. Accordingly, we extracted MMP and examined mechanisms of proMMP activation in whole tissue extracts of the adult rat myocardium. Incubation of extracts with serine proteases (i.e., trypsin or neutrophil elastase) at 37°C resulted in a time-dependent activation of proMMPs. Based on immunoblot and measurements of MMP activity by zymography, the molecular weight of active MMP was deduced to be 52 kDa. The second-order rate constant for activation of proMMP by serine protease was 5.5±0.2×105 M−1min−1 and for oxidized glutathione (GSSG) 1.5±0.1 M−1min−1. Incubation of the extract with both serine protease and GSSG increased the rate of activation 30-fold. Based on reverse zymographic analysis of collagenase inhibition, tissue inhibitors of metalloproteinases were identified. Indirect immunofluorescence localized proMMPs/MMPs to the endothelium and subendothelial space of the endocardium and throughout the interstitial space found between groups of muscle fibers. These results suggest that the mechanism of activation of MMPs by either a serine protease and by oxidizing, thiol-modifying reagents are mechanistically different and the presence of either a serine protease or GSSG synergistically increase the rate of activation of proMMPs. Our results also suggest that MMPs may be regulated by its own endogenous inhibitors. The contribution of this proteolytic enzyme to tissue remodeling and wound healing responses that occur in various diseases states remains to be established.

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Abbreviations

GSSG:

Oxidized Glutathione

MMP:

Matrix Metalloproteinase

NE:

Neutrophil Elastase

TIMP:

Tissue Inhibitor of Metalloproteinase

References

  1. Weber KT: Cardiac interstitium in health and disease: the fibrillar collagen network. J Am Coll Cardiol 13: 1637–1652, 1989

    PubMed  Google Scholar 

  2. Factor SM, Robinson TF: Comparative connective tissue structure-function relationships in biologic pumps. Lab Invest 58: 150–156, 1988

    PubMed  Google Scholar 

  3. Laurent GJ: Dynamic state of collagen: pathways of collagen degradationin vivo and their possible role in regulation of collagen mass. Am J Physiol 252: C1–9, 1987

    PubMed  Google Scholar 

  4. Factor SM, Robinson TF, Dominitz R, Cho SH: Alterations of the myocardial skeletal framework in acute myocardial infarction with an without ventricular rupture. Am J Cardiovasc Pathol 1: 91–97, 1987

    PubMed  Google Scholar 

  5. Chapman D, Weber KT, Eghbali M: Regulation of fibrillar collagen types I and III and basement membrane type IV collagen gene expression in pressure overloaded rat myocardium. Circ Res 67: 787–794, 1990

    PubMed  Google Scholar 

  6. Bishop JE, Greenbaum R, Gibson DG, Yacoub M, Laurent G: Enhanced deposition of predominantly type I collagen in myocardial disease. J Mol Cell Cardiol 22: 1157–1165, 1990

    PubMed  Google Scholar 

  7. Montfort I, Perez-Tamayo R: The distribution of collagenase in normal rat tissues. J Histochem Cytochem 23: 910–920, 1975

    PubMed  Google Scholar 

  8. Chakraborty A, Eghbali M: Collagenase activity in the normal rat myocardium: an immunohistochemical method. Histochemistry 92: 391–396, 1989

    PubMed  Google Scholar 

  9. Sato S, Ashraf M, Millard RW, Fujiwara H, Schwartz A: Connective tissue changes in early ischemia of porcine myocardium: an ultrastructural study. J Mol Cell Cardiol 15: 261–275, 1983

    PubMed  Google Scholar 

  10. Cannon RO, Butany JW, McManus BM, Speir E, Kravitz AB, Bolli R et al.: Early degradation of collagen after acute myocardial infarction in the rat. Am J Cardiol 52: 390–395, 1983

    PubMed  Google Scholar 

  11. Murphy G, Hembry RM, Hughes CE, Fosang AJ, Hardingham TE: Role and regulation of metalloproteinases in connective tissue turnover. Biochem Soc Trans 18: 812–815, 1990

    PubMed  Google Scholar 

  12. Chin J, Murphy G, Werb Z: Stromelysin, a connective tissue-degrading metalloendopeptidase secreted by stimulated rabbit synovial fibroblasts in parallel with collagenase. J Biol Chem 260: 12367–12376, 1985

    PubMed  Google Scholar 

  13. 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 166: 21–31, 1977

    PubMed  Google Scholar 

  14. Nagase H, Cawston TE, De Silva M, Barrett AJ: Identification of plasma kallikrein as an activator of latent collagenase in rheumatoid synovial fluid. Biochim Biophys Acta 702: 133–142, 1982

    PubMed  Google Scholar 

  15. Stricklin GP, Jeffrey JJ, Roswit WT, Eisen AZ: Human skin fibroblast procollagenase: mechanisms of activation by organomercurials and trypsin. Biochemistry 22: 61–68, 1983.

    PubMed  Google Scholar 

  16. Grant GA, Eisen AZ, Marmer BL, Roswit WT, Goldberg GI: The activation of human skin fibroblast procollagenase. Sequence identification of the major conversion products. J Biol Chem 262: 5886–5889, 1987

    PubMed  Google Scholar 

  17. Alexander CM, Werb Z: Proteinases and extracellular matrix re-modeling. Curr Opin Cell Biol 1: 974–982, 1989

    PubMed  Google Scholar 

  18. Takahashi S, Barry AC, Factor SM: Collagen degradation in ischaemic rat heart. Biochem J 265: 233–241, 1990

    PubMed  Google Scholar 

  19. Curello S, Ceconi C, Bigoli C, Ferrari R, Albertini A, Guarnieri C: Changes in the cardiac glutathione status after ischemia and reperfusion. Experientia 41: 42–43, 1985

    PubMed  Google Scholar 

  20. Caulfield JB, Wolkowicz P: Inducible collagenolytic activity in isolated, perfused rat hearts. Am J Pathol 131: 199–205, 1988

    PubMed  Google Scholar 

  21. Okada Y, Watanabe S, Nakanishi I, Kishi J, Hayakawa T, Watorek W et al.: Inactivation of tissue inhibitor of metalloproteinases by neutrophil elastase and other serine proteinases. FEBS Lett 229: 157–160, 1988

    PubMed  Google Scholar 

  22. Tyagi SC: Reversible inhibition of neutrophil elastase by thiol-modified alpha-1 protease inhibitor. J Biol Chem 266: 5279–5285, 1991

    PubMed  Google Scholar 

  23. Tyagi SC, Matsubara L, Weber KT: Direct extraction and estimation of collagenase(s) activity by zymography in microquantities of rat myocardium and uterus. Clin Biochem 26: 191–198, 1993

    PubMed  Google Scholar 

  24. Tyagi SC, Simon SR: Role of disulfide exchange in alpha-1 protease inhibitor. Biochemistry 1992: 10584–10590

  25. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254, 1976

    PubMed  Google Scholar 

  26. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970

    PubMed  Google Scholar 

  27. Granelli-Piperno A, Reich E: A study of protease and protease-inhibitor complexes in biological fluids. J Exp Med 148: 223–234, 1978

    PubMed  Google Scholar 

  28. Weber KT, Pick R, Janicki JS, Gadodia G, Lakier JB: Inadequate collagen tethers in dilated cardiopathy. Am Heart J 116: 1641–1646, 1988

    PubMed  Google Scholar 

  29. Brilla CG, Janicki JS, Weber KT: Impaired diastolic function and coronary reserve in genetic hypertension: role of interstitial fibrosis and medial thickening of intramyocardial coronary arteries. Circ Res 69: 107–115, 1991

    PubMed  Google Scholar 

  30. Okada Y, Nagase H, Harris ED Jr: A metalloproteinase from human rheumatoid synovial fibroblasts that digests connective tissue matrix components. Purification and characterization. J Biol Chem 261: 14245–14255, 1986

    PubMed  Google Scholar 

  31. Nagase H, Enghild JJ, Suzuki K, Salvesen G: Stepwise activation mechanisms of the precursor of matrix metalloproteinase 3 (stromelysin) by proteinases and (4-aminophenyl)mercuric acetate. Biochemistry 29: 5783–5789, 1990

    PubMed  Google Scholar 

  32. Suzuki K, Enghild JJ, Morodomi T, Salvesen G, Nagase H: Mechanisms anisms of activation of tissue procollagenase by matrix metalloproteinase 3 (stromelysin). Biochemistry 29: 10261–10270, 1990

    PubMed  Google Scholar 

  33. Hibbs MS, Hasty KA, Seyer JM, Kang AH, Mainardi CL: Biochemical and immunological characterization of the secreted forms of human neutrophil gelatinase. J Biol Chem 260: 2493–2500, 1985

    PubMed  Google Scholar 

  34. Hibbs MS, Hoidal JR, Kang AH: Expression of a metalloproteinase that degrades native type V collagen and denatured collagens by cultured human alveolar macrophages. J Clin Invest 80: 1644–1650, 1987

    PubMed  Google Scholar 

  35. Azzo W, Woessner JF Jr: Purification and characterization of an acid metalloproteinase from human articular cartilage. J Biol Chem 261: 5434–5441, 1986

    PubMed  Google Scholar 

  36. Woessner JF Jr, Taplin CJ: Purification and properties of a small latent matrix metalloproteinase of the rat uterus. J Biol Chem 263: 16918–16925, 1988

    PubMed  Google Scholar 

  37. Neurath H: Evolution of proteolytic enzymes. Science 224: 350–357, 1984

    PubMed  Google Scholar 

  38. Ishibashi M, Ito A, Sakyo K, Mori Y: Procollagenase activator produced by rabbit uterine cervical fibroblasts. Biochem J 241: 527–534, 1987

    PubMed  Google Scholar 

  39. Vater CA, Nagase H, Harris ED Jr: Purification of an endogenous activator of procollagenase from rabbit synovial fibroblast culture medium. J Biol Chem 258: 9374–9382, 1983

    PubMed  Google Scholar 

  40. Vaes G: Multiple steps in the activation of the inactive precursor of bone collagenase by trypsin. FEBS Lett 28: 198–200, 1972

    PubMed  Google Scholar 

  41. Springman EB, Angleton EL, Birkedal-Hansen H, Van Wart HE: Multiple modes of activation of latent human fibroblast collagenase: evidence for the role of a Cys73 active-site zinc complex in latency and a ‘cystein switch’ mechanism for activation. Proc Natl Acad Sci USA 87: 364–368, 1990

    PubMed  Google Scholar 

  42. Werb Z, Mainardi CL, Water CA, Harris ED Jr: Endogenous activation of latent collagenase by rheumatoid synovial cells. Evidence for a role of plasminogen activator. N Engl J Med 296: 1017–1023, 1977

    PubMed  Google Scholar 

  43. Mastrisian LM: Metalloproteinases and their inhibitors in matrix remodeling. Trends Genet 6: 121–125, 1990

    PubMed  Google Scholar 

  44. Reddi AH: Extracellular matrix and development. In: KA Piez, AH Reddi (eds) Extracellular matrix biochemistry. Elsevier, New York, 1984, pp 375–412

    Google Scholar 

  45. Grillo HC, Gross, J: Collagenolytic activity during mammalian wound repair. Dev Biol 15: 300–317, 1967

    PubMed  Google Scholar 

  46. Eisen AZ: Human skin collagenase: localization and distribution in normal human skin. J Invest Dermatol 52: 442–448, 1969

    PubMed  Google Scholar 

  47. Liotta LA, Steeg PS, Stetler-Stevenson WG: Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell 64: 327–336, 1991

    PubMed  Google Scholar 

  48. Cawston TE, Galloway WA, Mercer E, Murphy G, Reynolds JJ: Purification of rabbit bone inhibitor of collagenase. Biochem J 195: 159–165, 1981

    PubMed  Google Scholar 

  49. Dean DD, Woessner JF Jr: Extracts of human articular cartilage contain an inhibitor of tissue metalloproteinases. Biochem J 218: 277–280, 1984

    PubMed  Google Scholar 

  50. Campbell SE, Diaz-Arias AA, Weber KT: Ventricle rupture and fibrillar collagen degradation. Circ 84 (Suppl II): p2–232, 1991

    Google Scholar 

  51. Brilla CG, Campbell SE, Matsubara L, Weber KT: Advanced hypertensive heart disease in SHR: lisinopril-mediated regression of myocardial fibrosis. Circ 86 (Suppl I): I-329, 1992

    Google Scholar 

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Authors and Affiliations

  1. Division of Cardiology, Department of Internal Medicine, University of Missouri-Columbia, MA432 Medical Science Building, 65212, Columbia, Missouri, USA

    Suresh C. Tyagi, Anna Ratajska & Karl T. Weber

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  1. Suresh C. Tyagi
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  2. Anna Ratajska
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  3. Karl T. Weber
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Tyagi, S.C., Ratajska, A. & Weber, K.T. Myocardial matrix metalloproteinase(s): localization and activation. Mol Cell Biochem 126, 49–59 (1993). https://doi.org/10.1007/BF01772207

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  • DOI: https://doi.org/10.1007/BF01772207

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