Abstract
The heptahelical G protein-coupled receptors (GPCRs) receive and transmit a wide range of extracellular stimuli and induce a wide array of cellular responses by activating signaling kinases. It has become increasingly evident that the agonist-stimulated GPCR complexed with the adaptor protein, β-arrestin, serves as a focal point to recruit, activate, and target kinases to discrete subcellular compartments. This chapter describes a protocol to visualize the changes in the subcellular distribution of activated extracellular signal-regulated kinases 1 and 2 (ERK1/2) when induced by the angiotensin II type 1a receptor.
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
Lefkowitz, R. J. (2004) Historical review: a brief history and personal retrospective of seven-transmembrane receptors. Trends Pharmacol Sci 25, 413–22.
DeWire, S. M., Ahn, S., Lefkowitz, R. J., and Shenoy, S. K. (2007) Beta-arrestins and cell signaling. Annu Rev Physiol 69, 483–510.
Gesty-Palmer, D., and Luttrell, L. M. (2008) Heptahelical terpsichory. Who calls the tune? J Recept Signal Transduct Res 28, 39–58.
Ferguson, S. S. (2001) Evolving concepts in G protein-coupled receptor endocytosis: the role in receptor desensitization and signaling. Pharmacol Rev 53, 1–24.
Kendall, R. T., and Luttrell, L. M. (2009) Diversity in arrestin function. Cell Mol Life Sci 66, 2953–73.
Patel, P. A., Tilley, D. G., and Rockman, H. A. (2009) Physiologic and cardiac roles of beta-arrestins. J Mol Cell Cardiol 46, 300–8.
Beaulieu, J. M., and Caron, M. G. (2008) Looking at lithium: molecular moods and complex behaviour. Mol Interv 8, 230–41.
Gesty-Palmer, D., Flannery, P., Yuan, L., Corsino, L., Spurney, R., Lefkowitz, R. J., and Luttrell, L. M. (2009) A β-Arrestin–Biased Agonist of the Parathyroid Hormone Receptor (PTH1R) Promotes Bone Formation Independent of G Protein Activation. Science Translational Medicine 1, ra1.
Oakley, R. H., Laporte, S. A., Holt, J. A., Caron, M. G., and Barak, L. S. (2000) Differential affinities of visual arrestin, beta arrestin1, and beta arrestin2 for G protein-coupled receptors delineate two major classes of receptors. J Biol Chem 275, 17201–10.
Lefkowitz, R. J., and Shenoy, S. K. (2005) Transduction of receptor signals by beta-arrestins. Science 308, 512–7.
Shenoy, S. K. (2007) Seven-transmembrane receptors and ubiquitination. Circ Res 100, 1142–54.
Luttrell, L. M. (2002) Activation and targeting of mitogen-activated protein kinases by G-protein-coupled receptors. Can J Physiol Pharmacol 80, 375–82.
Shenoy, S. K., and Lefkowitz, R. J. (2005) Receptor-specific ubiquitination of beta-arrestin directs assembly and targeting of seven-transmembrane receptor signalosomes. J Biol Chem 280, 15315–24.
Shenoy, S. K., Barak, L. S., Xiao, K., Ahn, S., Berthouze, M., Shukla, A. K., Luttrell, L. M., and Lefkowitz, R. J. (2007) Ubiquitination of beta-arrestin links seven-transmembrane receptor endocytosis and ERK activation. J Biol Chem 282, 29549–62.
Acknowledgments
This work was supported by National Institutes of Health grant HL 080525.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Shenoy, S.K. (2011). Visualizing G Protein-Coupled Receptor Signalsomes Using Confocal Immunofluorescence Microscopy. In: Luttrell, L., Ferguson, S. (eds) Signal Transduction Protocols. Methods in Molecular Biology, vol 756. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-160-4_20
Download citation
DOI: https://doi.org/10.1007/978-1-61779-160-4_20
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-159-8
Online ISBN: 978-1-61779-160-4
eBook Packages: Springer Protocols