Split luciferase-based assay for simultaneous analyses of the ligand concentration- and time-dependent recruitment of β-arrestin2.
Bioluminescence
GPCR
Kinetic measurements
Ligand characterization
Split luciferase complementation
β-Arrestin recruitment
Journal
Analytical biochemistry
ISSN: 1096-0309
Titre abrégé: Anal Biochem
Pays: United States
ID NLM: 0370535
Informations de publication
Date de publication:
15 05 2019
15 05 2019
Historique:
received:
20
11
2018
revised:
22
02
2019
accepted:
25
02
2019
pubmed:
12
3
2019
medline:
26
12
2019
entrez:
12
3
2019
Statut:
ppublish
Résumé
Functional selectivity of agonists has gained increasing interest in G protein-coupled receptor (GPCR) research, e.g. due to expectations of drugs with reduced adverse effects. Different agonist-dependent GPCR conformations are conceived to selectively activate a balanced or imbalanced intracellular signalling response, involving e.g. different Gα subtypes, Gβγ-subunits and β-arrestins. To discriminate between the different signalling pathways (bias), sensitive techniques are needed that do not interfere with signalling. We applied split luciferase complementation to the GPCR/β-arrestin2 interaction and thoroughly analysed the influence of its implementation on intracellular signalling. This led to an assay enabling the functional characterization of ligands at the hH
Identifiants
pubmed: 30853375
pii: S0003-2697(18)31184-9
doi: 10.1016/j.ab.2019.02.023
pii:
doi:
Substances chimiques
Ligands
0
beta-Arrestin 2
0
Luciferases
EC 1.13.12.-
G-Protein-Coupled Receptor Kinase 2
EC 2.7.11.16
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
8-16Informations de copyright
Copyright © 2019. Published by Elsevier Inc.