Abstract
Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteinases associated with extracellular matrix degradation, cellular migration, tissue remodeling, and angiogenesis. The activity of MMPs is regulated by the tissue inhibitors of metalloproteinases (TIMPs). Zymography and reverse zymography are useful to detect MMPs and TIMPs activities from various samples, for example vitreous, retina, plasma, and so on. Sample proteins are separated in substrate containing polyacrylamide gel by electrophoresis. The gel is incubated and then stained with Coomassie Blue. MMPs’ activities are detected as clear bands.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Nagase, H., and Woessner, J. F. (1999) Matrix metalloproteinases. J. Biol. Chem. 274, 21491–21494.
Kon, C. H., Occleston, N. L., Charteris, D., Daniels, J., Aylward, G. W., and Khaw, P. T. (1998) A prospective study of matrix metalloproteinases in proliferative vitreoretinapathy. Invest. Ophthalmol. Vis. Sci. 39, 1524–1529.
Kvanta, A., Sarman, S., Fagerholm, P., Seregard, S., and Steen, B. (2000) Expression of matrix metalloproteinase-2 (MMP-2) and vascular endothelial growth factor (VEGF) in inflammation-associated corneal neovascularization. Exp. Eye Res. 70, 419–428.
Plantner, J. J., Jiang, C., and Smine, A. (1998) Increase in interphotoreceptor matrix gelatinase A (MMP-2) associated with age-related macular degeneration. Exp. Eye Res. 67, 637–645.
Chau, K. Y., Sivaprasad, S., Patel, N., Donaldson, T. A., Luthert, P. J., and Chong, N. V. (2008) Plasma levels of matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9) in age-related macular degeneration. Eye 22, 855–859.
Uemura, S., Matsushita, H., Li, W., Glassford, A. J., Asagami, T., Lee, K. H., Harrison, D. G., and Tsao P. S. (2001) Diabetes mellitus enhances vascular matrix metalloproteinase activity. Circ. Res. 88, 1291–1298.
Visse, R., and Nagase, H. (2003) Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry. Circ. Res. 92, 827–839.
Vu, T. H., Shipley, J. M., Bergers, G., Berger, J. E., Helms, J. A., Hanahan, D., Shapiro, S. D., Senior, R. M., and Werb, Z. (1998) MMP-9/gelatinase B is a key regulator of growth plate angiogenesis and apoptosis of hypertrophic chondrocytes. Cell 93, 411–422.
Bergers, G., Brekken, R., McMahon, G., Vu, T. H., Itoh, T., Tamaki, K., Tanzawa, K., Thorpe, P., Itohara, S., Werb, Z., and Hanahan, D. (2000) Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. Nat. Cell Biol. 2, 737–744.
Iwai, S., Aljada, A., Higa, A., Nakanishi-Ueda, T., Fukuda, S., Kamegawa, M., Iwabuchi, S., Ueda, T., Caballero, S., Browne, R., Afzal, A., Grant, M., Yasuhara, H., Koide, R., Oguchi, K., Dandona, P., and Armstrong, D. (2006) Activation of AP-1 and increased synthesis of MMP-9 in the rabbit retina induced by lipid hydroperoxide. Curr. Eye Res. 31, 337–346.
Brew, K., Dinakarpandian, D., and Nagase, H. (2000) Tissue inhibitors of metalloproteinases: evolution, structure and function. Biochem. Biophys. Acta. 1477, 267–283.
Nagase, H., Visse, R., and Murphy, G. (2006) Structure and function of matrix metalloproteinases and TIMPs. Cardiovasc. Res. 69, 562–573.
Snoek-van Beurden, P. A., and Von den Hoff, J. W. (2005) Zymographic techniques for the analysis of matrix metalloproteinases and their inhibitors. BioTechniques 38, 73–83.
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.
Fernández-Resa, P., Mira, E., Quesada, A. R. (1995) Enhanced detection of casein zymography of matrix metalloproteinases. Anal. Biochem. 224, 434–435.
Springman, E. B., Angleton, E. L., Birkedal-Hansen, H., and Van Wart, H. E. (1990) Multiple modes of activation of latent human fibroblast collagenase: evidence for the role of a Cys73 active-site zinc complex in latency and a “cysteine switch” mechanism for activation. Proc. Natl. Acad. Sci. USA 87, 364–368.
Okamoto, T., Akuta, T., Tamura, F., van Der Vliet, A., and Akaike, T. (2004) Molecular mechanism for activation and regulation of matrix metalloproteinases during bacterial infections and respiratory inflammation. Biol. Chem. 385, 997–1006.
Hawkes, S. P., Li, H., and Taniguchi, G. T. (2001) Zymography and reverse zymography for detecting MMPs, and TIMPs, in Matrix Metalloproteinase Protocols. (Clark, I. M., ed.), Humana, Totowa, NJ, pp. 399–410.
Woessner, J. F. Jr. (1995) Quantification of matrix metalloproteinases in tissue samples. Methods Enzymol. 248, 510–528.
Asano, Y., Iwai, S., Okazaki, M., Kumai, T., Munemasa, Y., Oonuma, S., Tadokoro, M., Kobayashi, S., and Oguchi, K. (2008) Matrix metalloproteinase-9 in spontaneously hypertensive hyperlipidemic rats. Pathophysiology In press.
Kamiya, Y., Iwai, S., Nara K., Okazaki, M., and Oguchi, K. (2005) Effects of green tea on matrix metalloproteinases in streptozotocin-induced diabetic rats. J. Clin. Biochem. Nutr. 37, 77–85.
Ogura, H., Nakanishi-Ueda, T., Ueda, T., Iwai, S., Uchida, S., Saito, Y., Taguchi, Y., Yasuhara, H., Armstrong, D., Oguchi, K., and Koide, R. (2007) Effect of a dihydrobenzofuran derivative on lipid hydroperoxide-induced rabbit corneal neovascularization. J. Pharmacol. Sci. 103, 234–240.
Vincenti, M. P. (2001) The matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) genes, in Matrix Metalloproteinase Protocols. (Clark, I. M., ed.), Humana, Totowa, NJ, pp. 121–148.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Iwai, S., Nakanishi-Ueda, T., Armstrong, D., Oguchi, K. (2008). Zymographical Techniques for Detectionof Matrix Metalloproteinases. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress I. Methods In Molecular Biology, vol 477. Humana Press. https://doi.org/10.1007/978-1-60327-517-0_10
Download citation
DOI: https://doi.org/10.1007/978-1-60327-517-0_10
Publisher Name: Humana Press
Print ISBN: 978-1-60327-218-6
Online ISBN: 978-1-60327-517-0
eBook Packages: Springer Protocols