Multidimensional Tracking of GPCR Signaling via Peroxidase-Catalyzed Proximity Labeling.

ABSTRACT: G-protein-coupled receptors (GPCRs) play critical roles in regulating physiological processes ranging from neurotransmission to cardiovascular function. Current methods for tracking GPCR signaling suffer from low throughput, modification or overexpression of effector proteins, and low temporal resolution. Here, we show that peroxidase-catalyzed proximity labeling can be combined with isobaric tagging and mass spectrometry to enable quantitative, time-resolved measurement of GPCR agonist response in living cells. Using this technique, termed "GPCR-APEX," we track activation and internalization of the angiotensin II type 1 receptor and the β2 adrenoceptor. These receptors co-localize with a variety of G proteins even before receptor activation, and activated receptors are largely sequestered

SUBMITTER: Paek J 

PROVIDER: S-EPMC5514552 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature