Summary
Matrix metalloproteinases (MMPs) are zinc-dependent proteases whose physiological roles include control of leukocyte migration. They are implicated in tissue destruction in inflammatory and infectious diseases. MMPs are not only capable of degrading all components of the extracellular matrix, but they also can modulate the immune response by cleaving cytokines and chemokines to alter their activity. Macrophages secrete a broad range of MMPs and represent a key source of MMPs in inflammatory lesions such as granulomas. Zymography is substrate-based gel electrophoresis that allows direct visualization of MMP activity. Here we describe measurement of MMP secretion from macrophages focusing on quantitative zymography. We also discuss complementary methods that should be used in parallel with zymography. The ability to analyze and quantify MMP secretion by macrophages offers an additional window through which to understand the contributions of macrophages to a wide variety of infectious, inflammatory, and immunologic disorders.
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Nagase, H., R. Visse, and G. Murphy. (2006). Structure and function of matrix metalloproteinases and TIMPs. Cardiovasc Res. 69: 562–573
Parks, W. C., C. L. Wilson, and Y. S. Lopez-Boado. (2004). Matrix metalloproteinases as modulators of inflammation and innate immunity. Nat Rev Immunol 4: 617–629
Brinckerhoff, C. E., and L. M. Matrisian. (2002). Matrix metalloproteinases: a tail of a frog that became a prince. Nat Rev Mol Cell Biol 3: 207–214
Elkington, P. T., M. O’Kane C, and J. S. Friedland. (2005).The paradox of matrix metalloproteinases in infectious disease. Clin Exp Immunol 142: 12–20
Brew, K., D. Dinakarpandian, and H. Nagase (2000). Tissue inhibitors of metalloproteinases: evolution, structure and function. Biochim Biophys Acta 1477: 267–283
Campbell, E. J., J. D. Cury, S. D. Shapiro, G. I. Goldberg, and H. G. Welgus. (1991). Neutral proteinases of human mononuclear phagocytes. Cellular differentiation markedly alters cell phenotype for serine proteinases, metalloproteinases, and tissue inhibitor of metalloproteinases. J Immunol 146: 1286–1293
Leber, T. M., and F. R. Balkwill. (1997). Zymography: a single-step staining method for quantitation of proteolytic activity on substrate gels. Anal Biochem 249: 24–28
Van Wart, H. E., and H. Birkedal-Hansen. (1990).The cysteine switch: a principle of regulation of metalloproteinase activity with potential applicability to the entire matrix metalloproteinase gene family. Proc Natl Acad Sci U S A 87: 5578–5582
Elkington, P. T., J. A. Green, and J. S. Friedland. (2006). Filter sterilization of highly infectious samples to prevent false negative analysis of matrix metalloproteinase activity. J Immunol Methods 309: 115–119
Acknowledgments
Some of the work described was supported by the Wellcome Trust (PE) and the MRC UK (JG)
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Elkington, P.T., Green, J.A., Friedland, J.S. (2009). Analysis of Matrix Metalloproteinase Secretion by Macrophages. In: Reiner, N. (eds) Macrophages and Dendritic Cells. Methods in Molecular Biology™, vol 531. Humana Press. https://doi.org/10.1007/978-1-59745-396-7_16
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DOI: https://doi.org/10.1007/978-1-59745-396-7_16
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