title: Kinetics and dynamics in the G protein-coupled receptor signaling cascade.
abstract: We describe optical and microscopy methods based on Förster resonance energy transfer, fluorescence recovery after photobleaching, and imaging cross-correlation spectroscopy that permit to determine kinetic and dynamic properties of key reactions involved G protein-coupled receptor (GPCR) signaling from the initial ligand binding step to the generation of the second messenger, cAMP. Well suited to determine rate-limiting reactions taking place along a GPCR signaling cascade in live cells, these techniques have also uncovered new concepts in GPCR signaling as well as many interesting mechanistic subtleties by which GPCRs transmit neurotransmitter and hormone signals into cells.
authors: Laboratory for GPCR Biology, Department of Pharmacology and Chemical Biology, University of Pittsburgh, School of Medicine, Pittsburgh, Pennsylvania, USA. jpv@pitt.edu
authors: Jean-Pierre
authors: JP
authors: Vilardaga
authors: Guillermo
authors: G
authors: Romero
authors: Timothy N
authors: TN
authors: Feinstein
authors: Vanessa L
authors: VL
authors: Wehbi
mesh_terms: D019390:Arrestins / Q000235:genetics / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D015870:Gene Expression; D057809:HEK293 Cells; D020962:Heterotrimeric GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D010281:Parathyroid Hormone / Q000378:metabolism*; D038761:Photobleaching; D011485:Protein Binding; D055550:Protein Stability; D018016:Receptors, Parathyroid Hormone / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics*
chemical_list: D019390:Arrestins; D008024:Ligands; D010281:Parathyroid Hormone; D018016:Receptors, Parathyroid Hormone; D000242:Cyclic AMP; D020962:Heterotrimeric GTP-Binding Proteins