Skip to main content

Analysis of Matrix Metalloproteinase Secretion by Macrophages

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 531))

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.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Nagase, H., R. Visse, and G. Murphy. (2006). Structure and function of matrix metalloproteinases and TIMPs. Cardiovasc Res. 69: 562–573

    Article  PubMed  CAS  Google Scholar 

  2. 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

    Article  PubMed  CAS  Google Scholar 

  3. 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

    Article  PubMed  CAS  Google Scholar 

  4. 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

    Article  PubMed  CAS  Google Scholar 

  5. Brew, K., D. Dinakarpandian, and H. Nagase (2000). Tissue inhibitors of metalloproteinases: evolution, structure and function. Biochim Biophys Acta 1477: 267–283

    Article  PubMed  CAS  Google Scholar 

  6. 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

    PubMed  CAS  Google Scholar 

  7. 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

    Article  PubMed  CAS  Google Scholar 

  8. 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

    Article  PubMed  CAS  Google Scholar 

  9. 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

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

Some of the work described was supported by the Wellcome Trust (PE) and the MRC UK (JG)

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jon S. Friedland .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2009 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

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

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-396-7_16

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-972-7

  • Online ISBN: 978-1-59745-396-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics