TY - JOUR
T1 - Conformational signatures in β-arrestin2 reveal natural biased agonism at a G-protein-coupled receptor
AU - Reyes-Alcaraz, Arfaxad
AU - Lee, Yoo Na
AU - Yun, Seongsik
AU - Hwang, Jong Ik
AU - Seong, Jae Young
N1 - Funding Information:
This work was supported by grants from the Research Program (NRF-2015M3A9E7029172) of the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT, and Future Planning. We want to thank especially to Professor Asuka Inoue from Tohoku University in Japan for the kind gift of HEK293 knockout cell lines used in this work.
Publisher Copyright:
© 2018, The Author(s).
Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2018/12/1
Y1 - 2018/12/1
N2 - Discovery of biased ligands and receptor mutants allows characterization of G-protein- and β-arrestin-mediated signaling mechanisms of G-protein-coupled receptors (GPCRs). However, the structural mechanisms underlying biased agonism remain unclear for many GPCRs. We show that while Galanin induces the activation of the galanin receptor 2 (Galr2) that leads to a robust stimulation toward Gαq-protein and β-arrestin1/2, an alternative ligand Spexin and its analog have biased agonism toward G-protein signaling relative to Galanin. We used intramolecular fluorescein arsenical hairpin bioluminescence resonance energy transfer-based biosensors of β-arrestin2 combined with NanoBit technology to measure β-arrestin2–Galr2 interactions in real-time living systems. We found that Spexin and Galanin induce specific active conformations of Galr2, which may lead to different internalization rates of the receptor as well as different signaling outputs. This work represents an additional pharmacological evidence of endogenous G-protein-biased agonism at a GPCR.
AB - Discovery of biased ligands and receptor mutants allows characterization of G-protein- and β-arrestin-mediated signaling mechanisms of G-protein-coupled receptors (GPCRs). However, the structural mechanisms underlying biased agonism remain unclear for many GPCRs. We show that while Galanin induces the activation of the galanin receptor 2 (Galr2) that leads to a robust stimulation toward Gαq-protein and β-arrestin1/2, an alternative ligand Spexin and its analog have biased agonism toward G-protein signaling relative to Galanin. We used intramolecular fluorescein arsenical hairpin bioluminescence resonance energy transfer-based biosensors of β-arrestin2 combined with NanoBit technology to measure β-arrestin2–Galr2 interactions in real-time living systems. We found that Spexin and Galanin induce specific active conformations of Galr2, which may lead to different internalization rates of the receptor as well as different signaling outputs. This work represents an additional pharmacological evidence of endogenous G-protein-biased agonism at a GPCR.
UR - http://www.scopus.com/inward/record.url?scp=85070417554&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85070417554&partnerID=8YFLogxK
U2 - 10.1038/s42003-018-0134-3
DO - 10.1038/s42003-018-0134-3
M3 - Article
C2 - 30272007
AN - SCOPUS:85070417554
VL - 1
JO - Communications Biology
JF - Communications Biology
SN - 2399-3642
IS - 1
M1 - 128
ER -