Fiche publication
Date publication
avril 2017
Journal
ChemMedChem
Auteurs
Membres identifiés du Cancéropôle Est :
Pr HIBERT Marcel
,
Dr VILLA Pascal
,
Dr BONNET Dominique
Tous les auteurs :
Valencia C, Dujet C, Margathe JF, Iturrioz X, Roux T, Trinquet E, Villa P, Hibert M, Dupuis E, Llorens-Cortes C, Bonnet D
Lien Pubmed
Résumé
Analogues of apelin-13 carrying diverse spacers and an ad hoc DY647-derived fluorophore were designed and synthesized by chemoselective acylation of α-hydrazinopeptides. The resulting probes retain very high affinity and efficacy for both the wild-type and SNAP-tagged apelin receptor (ApelinR). They give a time-resolved FRET (TR-FRET) signal with rare-earth lanthanides used as donor fluorophores grafted onto the SNAP-tagged receptor. This specific signal allowed the validation of a binding assay with a high signal-to-noise ratio. In such an assay, the most potent sub-nanomolar fluorescent probe was found to be competitively displaced by the endogenous apelin peptides with binding constants similar to those obtained in a classical radioligand assay. We have thus validated the first TR-FRET cell-based binding assay for ApelinR with potential high-throughput screening applications.
Mots clés
G protein-coupled receptors, TR-FRET assays, apelin, chemical ligation, fluorescent probes
Référence
ChemMedChem. 2017 Apr;:
