Visualizing G Protein-Coupled Receptor Signalsomes Using Confocal Immunofluorescence Microscopy
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.
Key wordsG protein-coupled receptor Confocal microscopy Extracellular signal-regulated kinase Arrestin Endocytosis Signaling Angiotensin G protein Scaffold
This work was supported by National Institutes of Health grant HL 080525.
- 8.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.Google Scholar