Abstract
Matrix metalloproteinases (MMPs) modulate cell–cell and cell–matrix interactions and the membrane-type MMPs (MT-MMPs) are especially important for pericellular proteolysis. Understanding the activity, regulation, and function of MT-MMPs would provide important insight for ongoing research in many diseases including cancer, fibrosis, and autoimmune diseases. In this chapter, we introduce the methods to express and purify recombinant MT-MMPs using Escherichia coli or mammalian cells, as well as how to detect the activity of MT-MMPs in proMMP-2 activation using gelatin zymography.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Nagase, H. and Woessner, J. F., Jr. (1999) Matrix metalloproteinases. J Biol Chem 274, 21491–21494.
Zucker, S., Pei, D., Cao, J., and Lopez-Otin, C. (2003) Membrane type-matrix metalloproteinases (MT-MMP). Curr Top Dev Biol 54, 1–74.
Nie, J. and Pei, D. (2004) Rapid inactivation of alpha-1-proteinase inhibitor by neutrophil specific leukolysin/membrane-type matrix metalloproteinase 6. Exp Cell Res 296, 145–150.
Nie, J. and Pei, D. (2003) Direct activation of pro-matrix metalloproteinase-2 by leukolysin/membrane-type 6 matrix metalloproteinase/matrix metalloproteinase 25 at the asn(109)-Tyr bond. Cancer Res 63, 6758–6762.
Jiang, A. and Pei, D. (2003) Distinct roles of catalytic and pexin-like domains in membrane-type matrix metalloproteinase (MMP)-mediated pro-MMP-2 activation and collagenolysis. J Biol Chem 278, 38765–38771.
Kang, T., Yi, J., Yang, W., Wang, X., Jiang, A., and Pei, D. (2000) Functional characterization of MT3-MMP in transfected MDCK cells: progelatinase a activation and tubulogenesis in 3-D collagen lattice. FASEB J 14, 2559–2568.
Wang, P., Wang, X., and Pei, D. (2004) Mint-3 regulates the retrieval of the internalized membrane-type matrix metalloproteinase, MT5-MMP, to the plasma membrane by binding to its carboxyl end motif EWV. J Biol Chem 279, 20461–20470.
Jiang, A., Lehti, K., Wang, X., Weiss, S. J., Keski-Oja, J., and Pei, D. (2001) Regulation of membrane-type matrix metalloproteinase 1 activity by dynamin-mediated endocytosis. Proc Natl Acad Sci U S A 98, 13693–13698.
Wang, X., Yi, J., Lei, J., and Pei, D. (1999) Expression, purification and characterization of recombinant mouse MT5-MMP protein products. FEBS Lett 462, 261–266.
Wang, P., Nie, J., and Pei, D. (2004) The hemopexin domain of membrane-type matrix metalloproteinase-1 (MT1-MMP) Is not required for its activation of proMMP2 on cell surface but is essential for MT1-MMP-mediated invasion in three-dimensional type I collagen. J Biol Chem 279, 51148–51155.
Sato, H., Takino, T., Kinoshita, T., Imai, K., Okada, Y., Stetler Stevenson, W. G., and Seiki, M. (1996) Cell surface binding and activation of gelatinase A induced by expression of membrane-type-1-matrix metalloproteinase (MT1-MMP). FEBS Lett 385, 238–240.
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
Nie, J., Pei, D. (2010). Expression and Purification of Membrane-Type MMPs. 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_6
Download citation
DOI: https://doi.org/10.1007/978-1-60327-299-5_6
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