S26-2 Subcellular Localization and Tracking of GPCRs with BRET

Program: Symposia
Session: S26-New Twists on GPCR Trafficking
Sunday, June 16, 2013: 9:00 AM-10:30 AM
Presentation Start Time: 9:30 AM
Room 206 (Moscone Center)
Nevin Lambert*
Georgia Health Sciences University, Augusta, GA
Talk Description:

Like other membrane proteins GPCRs transit multiple organelles and compartments between biogenesis and degradation. The subcellular location of GPCRs is highly regulated and important for their function, thus it is often necessary to determine the subcellular distribution of a particular receptor. We have found that bioluminescence resonance energy transfer (BRET) between membrane-targeted donors or acceptors and GPCRs tagged with fluorescent proteins or luciferases can be used to localize receptors and track their movement through subcellular compartments in living cells. We have used this method to monitor agonist-induced receptor internalization and recycling, and pharmacological chaperoning. The sensitivity of BRET allows receptors expressed at low levels to be localized with high spatial and temporal resolution. The method requires little user intervention for data acquisition or analysis, and is therefore suitable for large-scale experimental designs.

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