Development and Characterization of a Highly Selective Turn-On Fluorescent Ligand for ß3-Adrenergic Receptor.
Anal Chem
; 95(5): 2848-2856, 2023 02 07.
Article
em En
| MEDLINE
| ID: mdl-36700797
For the precise visualization of GPCR, subtype selectivity of turn-on fluorescent ligands is of major relevance. Although there are many thriving ß-adrenergic receptors (ß-ARs) probes, none of them are selective to the ß3-subtype, which severely limits the development of ß3-AR investigations. Using a polyethylene glycol (PEG) chain to conjugate the Py-5 fluorophore with mirabegron, we present here a highly selective fluorescent ligand, H2, for ß3-AR. It was established by the radioligand and NanoLuc-based bioluminescence resonance energy transfer (NanoBRET) binding experiments that molecule H2 has a substantially higher affinity for ß3-AR than the other two subtypes (1/3, 45-fold; 2/3, 16-fold). More crucially, when molecule H2 was incubated with ß3-AR, the turn-on fluorescent signals could be quickly released. The subsequent investigations, which included cell imaging, tissue imaging, and flow-cytometry analysis, proved that molecule H2 may make it possible to quickly and accurately fluorescently identify ß3-AR at different levels. We offer a prospective fluorescent turn-on ligand with exceptional selectivity for ß3-AR as a result of our combined efforts.
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Base de dados:
MEDLINE
Assunto principal:
Receptores Adrenérgicos beta
/
Agonistas Adrenérgicos beta
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article