Fractionation of Interphotoreceptor Matrix Metalloproteinases

  • James J. Plantner
  • Timothy A. Quinn
  • George J. Dwyer


Metalloproteinases (MPs) present in the interphotoreceptor matrix (IPM) may play a role in both the maintenance of normal retinal homeostasis and the progression of various degenerative retinal diseases. We wished to extend our previous studies of these enzymes and begin their characterization by first obtaining them in purified form.

IPM was obtained from fresh bovine eyes. The material was fractionated by a combination of gel filtration, DEAE ion exchange and gelatin- affinity chromatographies. Fractions were monitored by SDS-PAGE carried out on both plain gels and those preloaded with gelatin or casein (zymograms) and by quantitative analysis for enzyme activity Combination of the procedures described above provided a partial fractionation of the MPs as detected by zymography. A previously unrecognized 52 kDa caseinase was also revealed, possibly identical with MMP-3 (stromelysin).


Retinal Pigment Epithelium Trabecular Meshwork Proliferative Vitreoretinopathy Interphotoreceptor Retinoid Binding Protein Degenerative Retinal Disease 
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  1. 1.
    Hageman, G. and Johnson, L., 1991, Structure, composition and function of the retinal interphotoreceptor matrix, in Prog. Ret. Res., (N. Osborne and G.J. Chader, eds.), 10: 207–269.Google Scholar
  2. 2.
    Plantner, J.J., 1992, The presence of neutral metalloproteolytic activity and metalloproteinase inhibitors in the interphotoreceptor matrix, Curr. Eye Res. 11: 91–101.PubMedCrossRefGoogle Scholar
  3. 3.
    Plantner, J.J. and Drew, T.A., 1994, Polarized distribution of metalloproteinases in the bovine interphotoreceptor matrix, Exp. Eye Res. 59: 577–585.PubMedCrossRefGoogle Scholar
  4. 4.
    Plantner, J.J. and Quinn, T.A., Matrix Metalloproteinases in the Interphotoreceptor Matrix and Vitreous, submitted to Invest. Ophthalmol. Vis. Sci. for publication.Google Scholar
  5. 5.
    Jones, R.E., Moshyedi, P., Gallo, S., Tombran-Tink, J., Arand, G., Reid, D.A., Thompson, E.W., Chader, G.J. and Waldbillig, R.J., 1994, Characterization and novel activation of 72-kDa metalloproteinase in retinal interphotoreceptor matrix and Y-79 cell culture medium, Exp. Eye Res. 59: 257–269.PubMedCrossRefGoogle Scholar
  6. 6.
    Birkedal-Hansen, H., 1988, From tadpole collagenase to a family of matrix metalloproteinases, J. Oral Pathol. 17: 445–451.PubMedCrossRefGoogle Scholar
  7. 7.
    Matrisian, L.M., 1992, The matrix-degrading Metalloproteinases. BioEssays, 14: 455–463.PubMedCrossRefGoogle Scholar
  8. 8.
    Woessner, Jr., J.F., 1991, Matrix metalloproteinases and their inhibitors in connective tissue remodeling, FASEB J. 5: 2145–2154.PubMedGoogle Scholar
  9. 9.
    Kolkenbrock, H., Orgel, D., Hecker-Kia, A., Noack, W. and Ulbrich, N., 1991, The complex between a tissue inhibitor of metalloproteinases (TIMP-2) and 72-kDa progelatinase is a metalloproteinase inhibitor, Eur. J. Biochem. 198: 775–781.PubMedCrossRefGoogle Scholar
  10. 10.
    Moscatelli, D. and Rifkin, D.B., 1988, Membrane and matrix localization of proteinases: a common theme in tumor cell invasion and angiogenesis, Biochim. Biophys. Acta 948: 67–85.PubMedGoogle Scholar
  11. 11.
    Fini, M.E., Yue, B.Y.J.T. and Sugar, J., 1992, Collagenolytic/Gelatinolytic Metalloproteinases in Normal and Keratoconus Corneas, Curr. Eye Res. 11: 849–862.PubMedCrossRefGoogle Scholar
  12. 12.
    Alexander, J.P., Samples, J.R., Van Buskirk, E.M. and Acott, T.S., 1991, Expression of Matrix Metalloproteinases and Inhibitor by Human Trabecular Meshwork, Invest Ophthalmol Vis Sci. 32: 172–180.PubMedGoogle Scholar
  13. 13.
    Alexander, J.P., Bradley, J.M., Gabourel, J.D. and Acott, T.S., 1990, Expression of Matrix Metalloproteinases and Inhibitor by Human Retinal Pigment Epithelium, Invest Ophthalmol. Vis. Sci. 31: 2520–2528.PubMedGoogle Scholar
  14. 14.
    Hunt, R.C., Fox, A., Al Pakalnis, V, Sigel, M.M., Kosnosky, W., Choudhury, P. and Black, E.P, 1993, Cytokines Cause Cultured Retinal Pigment Epithelial Cells to Secrete Metalloproteinases and to Contract Collagen Gels, Invest. Ophthalmol. Vis. Sci. 34: 3179–3186.PubMedGoogle Scholar
  15. 15.
    Wentworth, J.S., Paterson, C.A. and Gray, R.D., 1992, Effect of a Metalloproteinase Inhibitor on Established Corneal Ulcers After an Alkali Burn, Invest. Ophthalmol. Vis. Sci. 33: 2174–2179.PubMedGoogle Scholar
  16. 16.
    Patz, A., 1980, Studies on retinal neovascularization, Invest. Ophthalmol. Vis. Sci. 19: 1133–1138.PubMedGoogle Scholar
  17. 17.
    Machemer, R., 1988, Proliferative vitreoretinopathy (PVR): A personal account of its pathogenesis and treatment, Invest. Ophthalmol. Vis. Sci. 29: 1771–1783.PubMedGoogle Scholar
  18. 18.
    Green, W.R., McDonnell, P.J. and Yeo, J.H., 1985, Pathologic features of senile macular degeneration, Ophthalmol. 92: 615–627.CrossRefGoogle Scholar
  19. 19.
    Taylor, C.M., Weiss, J.B., Kissun, R.D., and Garner, A., 1986, Effect of oxygen tension on the quantities of procollagenase-activating angiogenesis factor in the vitreous humour of oxygen treated kittens, Brit. J. Ophthalmol 70: 162–168.CrossRefGoogle Scholar
  20. 20.
    Caldwell, R.B., Slapnick, S.M., and Roque, R.S., 1989, RPE-associated extracellular matrix changes accompany retinal vascular proliferation and retino-vitreal membranes in a new model for proliferative retinopathy: the dystrophic rat, In “ Inherited and Environmentally Induced Retinal Degenerations,” Alan R. Liss, Pp. 393–407.Google Scholar
  21. 21.
    Jones, S.E., Jomary, C. and Neal, M. J., 1994, Expression of TIMP3 mRNA is elevated in Retinas Affected by Simplex Retinitis Pigmentosa, FEBS Lett. 352: 171–174.PubMedCrossRefGoogle Scholar
  22. 22.
    Weber, B.H.F., Vogt, G. Pruett, R.C., Stohr, H. and Felbor, U., 1994, Mutations in the Tissue Inhibitor of Metalloproteinases-3 (TIMP3) in Patients with Sorsby’ s Fundus Dystrophy, Nature Genetics 8: 352–356.PubMedCrossRefGoogle Scholar
  23. 23.
    Adler, A.J. and Severin, K.M., 1981, Proteins of the Bovine Interphotoreceptor Matrix: Tissues of Origin, Exp.Eye Res. 32: 155–769.CrossRefGoogle Scholar
  24. 24.
    Plantner, J.J., 1992, High Molecular Weight Mucin-like Glycoproteins of the Bovine Interphotoreceptor Matrix, Exp. Eye Res. 54: 113–125.PubMedCrossRefGoogle Scholar
  25. 25.
    Heussen, C. and Dowdle, E.B., 1980, Electrophoretic Analysis of Plasminogen Activators in Polyacrylamide Gels Containing Sodium Dodecyl Sulfate and Copolymerized Substrates, Anal. Biochem. 102: 196–202.PubMedCrossRefGoogle Scholar
  26. 26.
    Plantner, J.J., 1991, Amicroassay for proteolytic activity, Anal. Biochem. 195: 129–131.PubMedCrossRefGoogle Scholar
  27. 27.
    Weingarten, H., Martin, R. and Feder, J., 1985, Synthetic Substrates of Vertebrate Collagenase, Biochem, 24: 6730–6734.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • James J. Plantner
    • 1
  • Timothy A. Quinn
    • 1
  • George J. Dwyer
    • 1
  1. 1.Department of OphthalmologyCase Western Reserve UniversityClevelandUSA

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