Monitoring G protein-coupled receptor and β-arrestin trafficking in live cells using enhanced bystander BRET

Yoon Namkung, Christian Le Gouill, Viktoria Lukashova, Hiroyuki Kobayashi, Mireille Hogue, Etienne Khoury, Mideum Song, Michel Bouvier () and Stéphane A. Laporte ()
Additional contact information
Yoon Namkung: Research Institute of the McGill University Health Center (RI-MUHC), McGill University
Christian Le Gouill: Université de Montréal
Viktoria Lukashova: Université de Montréal
Hiroyuki Kobayashi: Université de Montréal
Mireille Hogue: Université de Montréal
Etienne Khoury: Research Institute of the McGill University Health Center (RI-MUHC), McGill University
Mideum Song: Research Institute of the McGill University Health Center (RI-MUHC), McGill University
Michel Bouvier: Université de Montréal
Stéphane A. Laporte: Research Institute of the McGill University Health Center (RI-MUHC), McGill University

Nature Communications, 2016, vol. 7, issue 1, 1-12

Abstract: Abstract Endocytosis and intracellular trafficking of receptors are pivotal to maintain physiological functions and drug action; however, robust quantitative approaches are lacking to study such processes in live cells. Here we present new bioluminescence resonance energy transfer (BRET) sensors to quantitatively monitor G protein-coupled receptors (GPCRs) and β-arrestin trafficking. These sensors are based on bystander BRET and use the naturally interacting chromophores luciferase (RLuc) and green fluorescent protein (rGFP) from Renilla. The versatility and robustness of this approach are exemplified by anchoring rGFP at the plasma membrane or in endosomes to generate high dynamic spectrometric BRET signals on ligand-promoted recruitment or sequestration of RLuc-tagged proteins to, or from, specific cell compartments, as well as sensitive subcellular BRET imaging for protein translocation visualization. These sensors are scalable to high-throughput formats and allow quantitative pharmacological studies of GPCR trafficking in real time, in live cells, revealing ligand-dependent biased trafficking of receptor/β-arrestin complexes.

Date: 2016
References: Add references at CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/ncomms12178 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms12178

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/ncomms12178

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().