Ligand-directed covalent labelling of a GPCR with a fluorescent tag in live cells.
Commun Biol
; 3(1): 722, 2020 11 27.
Article
in En
| MEDLINE
| ID: mdl-33247190
ABSTRACT
To study the localisation of G protein-coupled receptors (GPCR) in their native cellular environment requires their visualisation through fluorescent labelling. To overcome the requirement for genetic modification of the receptor or the limitations of dissociable fluorescent ligands, here we describe rational design of a compound that covalently and selectively labels a GPCR in living cells with a fluorescent moiety. We designed a fluorescent antagonist, in which the linker incorporated between pharmacophore (ZM241385) and fluorophore (sulfo-cyanine5) is able to facilitate covalent linking of the fluorophore to the adenosine A2A receptor. We pharmacologically and biochemically demonstrate irreversible fluorescent labelling without impeding access to the orthosteric binding site and demonstrate its use in endogenously expressing systems. This offers a non-invasive and selective approach to study function and localisation of native GPCRs.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Triazines
/
Triazoles
/
Receptors, G-Protein-Coupled
/
Fluorescent Dyes
Limits:
Humans
Language:
En
Journal:
Commun Biol
Year:
2020
Document type:
Article
Affiliation country: