Abstract
All ligands of the epidermal growth factor receptor (EGFR) are made as membrane anchored precursors that can be proteolytically processed and released from the plasma membrane. This process, which is referred to as protein ectodomain shedding, is emerging as a key regulator of the function of EGFR ligands. In light of the important roles of EGFR signaling in development and disease, it will be important to understand more about the regulation of proteolytic processing of EGFR ligands. This chapter describes a sensitive and semiquantitative method to measure ectodomain shedding of EGFR ligands that was designed to facilitate studies of this process in cells.
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
Yarden Y. and Sliwkowski M. X. (2001) Untangling the ErbB signaling network.Nat. Rev. Mol. Cell. Biol. 2, 127–137.
Gschwind A., Fischer O. M., and Ullrich A. (2004) The discovery of receptortyrosine kinases: targets for cancer therapy. Nat. Rev. Cancer 4, 361–370.
Schlessinger J. (2002) Ligand-induced, receptor-mediated dimerization and activationof EGF receptor. Cell 110, 669–672.
Burgess A. W., Cho H. S., Eigenbrot C., et al. (2003) An open-and-shut case?Recent insights into the activation of EGF/ErbB receptors. Mol. Cell. 12, 541–552.
Harris R. C., Chung, E., and Coffey R. J. (2003) EGF receptor ligands. Exp.Cell. Res. 284, 2–13.
Brachmann R., Lindquist P. B., Nagashima M., et al. (1989) TransmembraneTGF-alpha precursors activate EGF/TGF-alpha receptors. Cell 56, 691–700.
Wong S. T., Winchell L. F., McCune B. K., et al. (1989) The TGF-alpha precursorexpressed on the cell surface binds to the EGF receptor on adjacent cells,leading to signal transduction. Cell 56, 495–506.
Blobel C. P. (2004) ADAMs: key players in EGFR-signaling, development anddisease. Nat. Rev. Mol. Cell Biol. 6, 32–43.
Dong J., Opresko L. K., Dempsey P. J., Lauffenburger D. A., Coffey R. J., andWiley H. S. (1999) Metalloprotease-mediated ligand release regulates autocrinesignaling through the epidermal growth factor receptor. Proc. Natl. Acad. Sci.USA 96, 6235–6240.
Prenzel N., Zwick E., Daub H., et al. (1999) EGF receptor transactivation byG-protein-coupled receptors requires metalloproteinase cleavage of proHB-EGF. Nature 402, 884–888.
Schafer B., Gschwind, A., and Ullrich A. (2004) Multiple G-protein-coupledreceptor signals converge on the epidermal growth factor receptor to promotemigration and invasion. Oncogene 23, 991–999.
Peschon J. J., Slack J. L., Reddy P., et al. (1998) An essential role for ectodomainshedding in mammalian development. Science 282, 1281–1284.
Jackson L. F., Qiu T. H., Sunnarborg S. W., et al. (2003) Defective valvulogenesisin HB-EGF and TACE-null mice is associated with aberrant BMP signaling.EMBO J. 22, 2704–2716.
Sahin U., Weskamp G., Zhou H. M., et al. (2004) Distinct roles for ADAM10and ADAM17 in ectodomain shedding of six EGFR-ligands. J. Cell. Biol. 164, 769–779.
Zheng Y., Schlondorff, J., and Blobel C. P. (2002) Evidence for regulation of thetumor necrosis factor alpha-convertase (TACE) by protein-tyrosine phosphatasePTPH1. J. Biol. Chem. 277, 42,463–42,470.
Hartmann D., de Strooper B., Serneels L., et al. (2002) The disintegrin/metalloproteaseADAM 10 is essential for Notch signaling but not for alpha-secretaseactivity in fibroblasts. Hum. Mol. Genet. 11, 2615–2624.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Sahin, U., Weskamp, G., Zheng, Y., Chesneau, V., Horiuchi, K., Blobel, C.P. (2006). A Sensitive Method to Monitor Ectodomain Shedding of Ligands of the Epidermal Growth Factor Receptor. In: Patel, T.B., Bertics, P.J. (eds) Epidermal Growth Factor. Methods in Molecular Biology, vol 327. Humana Press. https://doi.org/10.1385/1-59745-012-X:99
Download citation
DOI: https://doi.org/10.1385/1-59745-012-X:99
Publisher Name: Humana Press
Print ISBN: 978-1-58829-421-0
Online ISBN: 978-1-59745-012-6
eBook Packages: Springer Protocols