Using the novel HiBiT tag to label cell surface relaxin receptors for BRET proximity analysis.
Bioluminescence resonance energy transfer
G protein‐coupled receptor
HiBiT
Nanoluc
RXFP1
Relaxin
Journal
Pharmacology research & perspectives
ISSN: 2052-1707
Titre abrégé: Pharmacol Res Perspect
Pays: United States
ID NLM: 101626369
Informations de publication
Date de publication:
08 2019
08 2019
Historique:
received:
25
01
2019
revised:
11
07
2019
accepted:
16
07
2019
entrez:
7
8
2019
pubmed:
7
8
2019
medline:
30
4
2020
Statut:
epublish
Résumé
Relaxin family peptide 1 (RXFP1) is the receptor for relaxin a peptide hormone with important therapeutic potential. Like many G protein-coupled receptors (GPCRs), RXFP1 has been reported to form homodimers. Given the complex activation mechanism of RXFP1 by relaxin, we wondered whether homodimerization may be explicitly required for receptor activation, and therefore sought to determine if there is any relaxin-dependent change in RXFP1 proximity at the cell surface. Bioluminescence resonance energy transfer (BRET) between recombinantly tagged receptors is often used in GPCR proximity studies. RXFP1 targets poorly to the cell surface when overexpressed in cell lines, with the majority of the receptor proteins sequestered within the cell. Thus, any relaxin-induced changes in RXFP1 proximity at the cell surface may be obscured by BRET signal originating from intracellular compartments. We therefore, utilized the newly developed split luciferase system called HiBiT to specifically label the extracellular terminus of cell surface RXFP1 receptors in combination with mCitrine-tagged receptors, using the GABA
Identifiants
pubmed: 31384473
doi: 10.1002/prp2.513
pii: PRP2513
pmc: PMC6667744
doi:
Substances chimiques
RXFP1 protein, human
0
Receptors, G-Protein-Coupled
0
Receptors, Peptide
0
Relaxin
9002-69-1
Luciferases
EC 1.13.12.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e00513Déclaration de conflit d'intérêts
The authors declare no competing interests.
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