Article (Scientific journals)
A dynamic and screening-compatible nanoluciferase-based complementation assay enables profiling of individual GPCR-G protein interactions
Laschet, Céline; Dupuis, Nadine; Hanson, Julien
2019In Journal of Biological Chemistry, 294 (11), p. 4079-4090
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Keywords :
G protein-coupled receptor (GPCR); G protein, high-throughput screening (HTS); drug screening; pharmacology; functional selectivity; biased signaling; complementation assay; dopamine; Nanoluciferase
Abstract :
[en] G protein-coupled receptors (GPCRs) are currently the target of more than 30% of the marketed medicines. However, there is an important medical need for ligands with improved pharmacological activities on validated drug targets. Moreover, most of these ligands remain poorly characterized, notably because of a lack of pharmacological tools. Thus, there is an important demand for innovative assays that can detect and drive the design of compounds with novel or improved pharmacological properties. In particular, a functional and screening-compatible GPCR-G protein interaction assay is still unavailable. Here, we report on a nanoluciferase-based complementation technique to detect ligands that promote a GPCR-G protein interaction. We demonstrate that our system can be used to profile compounds with regard to the G proteins they activate through a given GPCR. Furthermore, we established a proof of applicability of screening for distinct G proteins on dopamine receptor D2 whose differential coupling to Gαi/o family members has been extensively studied. In a D2-Gαi1 versus D2- Gαo screening, we retrieved five agonists that are currently being used in antiparkinsonian medications. We determined that in this assay, piribedil and pergolide are full agonists for the recruitment of Gαi1 but are partial agonists for Gαo, that the agonist activity of ropinirole is biased in favor of Gαi1 recruitment, and that the agonist activity of apomorphine is biased for Gαo. We proposed that this newly developed assay could be used to develop molecules that selectively modulate a particular G protein pathway.
Research center :
Giga-Signal Transduction - ULiège
Centre Interfacultaire de Recherche du Médicament - CIRM
Disciplines :
Life sciences: Multidisciplinary, general & others
Pharmacy, pharmacology & toxicology
Biochemistry, biophysics & molecular biology
Author, co-author :
Laschet, Céline ;  Université de Liège - ULiège > Département de pharmacie > Chimie pharmaceutique
Dupuis, Nadine
Hanson, Julien  ;  Université de Liège - ULiège > Département de pharmacie > Chimie pharmaceutique
Language :
Title :
A dynamic and screening-compatible nanoluciferase-based complementation assay enables profiling of individual GPCR-G protein interactions
Publication date :
15 March 2019
Journal title :
Journal of Biological Chemistry
Publisher :
American Society for Biochemistry and Molecular Biology, United States - Maryland
Volume :
Issue :
Pages :
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
F.R.S.-FNRS - Fonds de la Recherche Scientifique [BE]
FWB - Fédération Wallonie-Bruxelles [BE]
Fonds Léon Fredericq [BE]
FRIA - Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture [BE]
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