Conformation- and activation-based BRET sensors differentially report on GPCR-G protein coupling.
Sci Signal
; 17(841): eadi4747, 2024 06 18.
Article
de En
| MEDLINE
| ID: mdl-38889226
ABSTRACT
G protein-coupled receptors (GPCRs) regulate cellular signaling processes by coupling to diverse combinations of heterotrimeric G proteins composed of Gα, Gß, and Gγ subunits. Biosensors based on bioluminescence resonance energy transfer (BRET) have advanced our understanding of GPCR functional selectivity. Some BRET biosensors monitor ligand-induced conformational changes in the receptor or G proteins, whereas others monitor the recruitment of downstream effectors to sites of G protein activation. Here, we compared the ability of conformation-and activation-based BRET biosensors to assess the coupling of various class A and B GPCRs to specific Gα proteins in cultured cells. These GPCRs included serotonin 5-HT2A and 5-HT7 receptors, the GLP-1 receptor (GLP-1R), and the M3 muscarinic receptor. We observed different signaling profiles between the two types of sensors, highlighting how data interpretation could be affected by the nature of the biosensor. We also found that the identity of the Gßγ subunits used in the assay could differentially influence the selectivity of a receptor toward Gα subtypes, emphasizing the importance of the receptor-Gßγ pairing in determining Gα coupling specificity. Last, the addition of epitope tags to the receptor could affect stoichiometry and coupling selectivity and yield artifactual findings. These results highlight the need for careful sensor selection and experimental design when probing GPCR-G protein coupling.
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Techniques de biocapteur
/
Récepteurs couplés aux protéines G
/
Techniques de transfert d'énergie par résonance de bioluminescence
Limites:
Humans
Langue:
En
Journal:
Sci Signal
Sujet du journal:
CIENCIA
/
FISIOLOGIA
Année:
2024
Type de document:
Article
Pays d'affiliation:
Canada