Abstract
A number of matrix metalloproteinases (MMPs) and their endogenous inhibitors called tissue inhibitors of metalloproteinases (TIMPs) from natural sources have been identified. This chapter describes the purification methods of MMPs-1, -2, -3, -7, -8, -9, -10, -12, and -13 and TIMPs-1 and -2. The sources of the proteins and assay methods to detect their activities are also described.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Massova, I., Kotra, L. P., Fridman, R., and Mobashery, S. (1998) Matrix metalloproteinases – structures, evolution, and diversification. FASEB J 12, 1075–1095.
Gomez, D. E., Alonso, D. F., Yoshiji, H., and Thorgeirsson, U. P. (1997) Tissue inhibitors of metalloproteinases—structure, regulation and biological functions. Eur J Cell Biol 74, 111–122.
Imai, K., Yokohama, Y., Nakanishi, I., Ohuchi, E., Fujii, Y., Nakai, N., and Okada, Y. (1995) Matrix metalloproteinase 7 (matrilysin) from human rectal carcinoma cells. Activation of the precursor, interaction with other matrix metalloproteinases and enzymic properties. J Biol Chem 270, 6691–6697.
Woessner, J. F., Jr. and Taplin, C. J. (1988) Purification and properties of a small latent matrix metalloproteinase of the rat uterus. J Biol Chem 263, 16918–16925.
Morodomi, T., Ogata, Y., Sasaguri, Y., Morimatsu, M., and Nagase, H. (1992) Purification and characterization of matrix metalloproteinase 9 from U937 monocytic leukemia and HT1080 fibrosarcoma cells. Biochem J 285, 603–611.
Windsor, L. J., Grenett, H., Birkedal-Hansen, B., Bodden, M. K., Engler, J. A., and Birkedal-Hansen, H. (1993) Cell type-specific regulation of SL-1 and SL-2 genes. Induction of the SL-2 gene but not the SL-1 gene by human keratinocytes in response to cytokines and phorbolesters. J Biol Chem 268, 17341–17347.
Nakamura, H., Fujii, Y., Ohuchi, E., Yamamoto, E., and Okada, Y. (1998) Activation of the precursor of human stromelysin 2 and its interactions with other matrix metalloproteinases. Eur J Biochem 253, 67–75.
Knight, C. G., Willenbrock, F., and Murphy, G. (1992) A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases. FEBS Lett 296, 263–266.
Cawston, T. E. and Barrett, A. J. (1979) A rapid and reproducible assay for collagenase using [1-14C]acetylated collagen. Anal Biochem 99, 340–345.
Suzuki, K., Enghild, J. J., Morodomi, T., Salvesen, G., and Nagase, H. (1990) Mechanisms of activation of tissue procollagenase by matrix metalloproteinase 3 (stromelysin). Biochemistry 29, 10261–10270.
Itoh, Y., Binner, S., and Nagase, H. (1995) Steps involved in activation of the complex of pro-matrix metalloproteinase 2 (progelatinase A) and tissue inhibitor of metalloproteinases (TIMP)-2 by 4-aminophenylmercuric acetate. Biochem J 308, 645–651.
Ward, R. V., Hembry, R. M., Reynolds, J. J., and Murphy, G. (1991) The purification of tissue inhibitor of metalloproteinases-2 from its 72 kDa progelatinase complex. Demonstration of the biochemical similarities of tissue inhibitor of metalloproteinases-2 and tissue inhibitor of metalloproteinases-1. Biochem J, 278, 179–187.
Gunja-Smith, Z., Nagase, H., and Woessner, J. F., Jr. (1989) Purification of the neutral proteoglycan-degrading metalloproteinase from human articular cartilage tissue and its identification as stromelysin matrix metalloproteinase-3. Biochem J 258, 115–119.
Nagase, H. (1995) Human stromelysins 1 and 2. Methods Enzymol 248, 449–470.
Nagase, H., Fields, C. G., and Fields, G. B. (1994) Design and characterization of a fluorogenic substrate selectively hydrolyzed by stromelysin 1 (matrix metalloproteinase- 3). J Biol Chem 269, 20952–20957.
Ito, A. and Nagase, H. (1988) Evidence that human rheumatoid synovial matrix metalloproteinase 3 is an endogenous activator of procollagenase. Arch Biochem Biophys 267, 211–216.
Engelbrecht, S., Pieper, E., Macartney, H. W., Rautenberg, W., Wenzel, H. R., and Tschesche, H. (1982) Separation of the human leucocyte enzymes alanine aminopeptidase, cathepsin G, collagenase, elastase and myeloperoxidase. Hoppe Seylers Z Physiol Chem 363, 305–315.
Knäuper, V., Kramer, S., Reinke, H., and Tschesche, H. (1990) Characterization and activation of procollagenase from human polymorphonuclear leucocytes. N-terminal sequence determination of the proenzyme and various proteolytically activated forms. Eur J Biochem 189, 295–300.
Knäuper, V., Wilhelm, S. M., Seperack, P. K., DeClerck, Y. A., Langley, K. E., Osthues, A., and Tschesche, H. (1993) Direct activation of human neutrophil procollagenase by recombinant stromelysin. Biochem J 295, 581–586.
Moore, W. M. and Spilburg, C. A. (1986) Purification of human collagenases with a hydroxamic acid affinity column. Biochemistry 25, 5189–5195.
Banda, M. J. and Werb, Z. (1981) Mouse macrophage elastase. Purification and characterization as a metalloproteinase. Biochem J 193, 589–605.
Sudbeck, B. D., Jeffrey, J. J., Welgus, H. G., Mecham, R. P., McCourt, D., and Parks, W. C. (1992) Purification and characterization of bovine interstitial collagenase and tissue inhibitor of metalloproteinases. Arch Biochem Biophys 293, 370–376.
Cawston, T. E., Noble, D. N., Murphy, G., Smith, A. J., Woodley, C., and Hazleman, B. (1986) Rapid purification of tissue inhibitor of metalloproteinases from human plasma and identification as a gamma-serum protein. Biochem J 238, 677–682.
Umenishi, F., Umeda, M., and Miyazaki, K. (1991) Efficient purification of TIMP-2 from culture medium conditioned by human hepatoma cell line, and its inhibitory effects on metalloproteinases and in vitro tumor invasion. J Biochem (Tokyo) 110, 189–195.
Okada, Y., Nagase, H., and Harris, E. D., Jr. (1986) A metalloproteinase from human rheumatoid synovial fibroblasts that digests connective tissue matrix components. Purification and characterization. J Biol Chem 261, 14245–14255.
Lark, M. W., Walakovits, L. A., Shah, T. K., Vanmiddlesworth, J., Cameron, P. M., and Lin, T. Y. (1990) Production and purification of prostromelysin and procollagenase from IL-1 β-stimulated human gingival fibroblasts. Connect Tissue Res 25, 49–65.
Ogata, Y., Itoh, Y., and Nagase, H. (1995) Steps involved in activation of the pro-matrix metalloproteinase 9 (progelatinase B)-tissue inhibitor of metallopro-teinases-1 complex by 4-aminophenylmercuric acetate and proteinases. J Biol Chem 270, 18506–18511.
Shapiro, S. D., Kobayashi, D. K., and Ley, T. J. (1993): Cloning and characterization of a unique elastolytic metalloproteinase produced by human alveolar macrophages. J Biol Chem 268, 23824–23829
Acknowledgments
We thank Linda Chung for useful comments. This work is supported by NIH grants AR39189 and AR40994.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Shimokawa, Ki., Nagase, H. (2010). Purification of MMPs and TIMPs. In: Clark, I. (eds) Matrix Metalloproteinase Protocols. Methods in Molecular Biology, vol 622. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-299-5_8
Download citation
DOI: https://doi.org/10.1007/978-1-60327-299-5_8
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-298-8
Online ISBN: 978-1-60327-299-5
eBook Packages: Springer Protocols