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

Published

Journal Article

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 from G proteins upon internalization. Additionally, the two receptors show differing internalization kinetics, and we identify the membrane protein LMBRD2 as a potential regulator of β2 adrenoceptor signaling, underscoring the value of a dynamic view of receptor function.

Full Text

Duke Authors

Cited Authors

  • Paek, J; Kalocsay, M; Staus, DP; Wingler, L; Pascolutti, R; Paulo, JA; Gygi, SP; Kruse, AC

Published Date

  • April 6, 2017

Published In

Volume / Issue

  • 169 / 2

Start / End Page

  • 338 - 349.e11

PubMed ID

  • 28388415

Pubmed Central ID

  • 28388415

Electronic International Standard Serial Number (EISSN)

  • 1097-4172

Digital Object Identifier (DOI)

  • 10.1016/j.cell.2017.03.028

Language

  • eng

Conference Location

  • United States