Click-Functionalized Cyanine Fluorogenic Dimers for Improved Detection of GPCRs: Application to imaging of ApelinR in Living Cells.

Fiche publication

Date publication

avril 2025

Journal

Chemistry (Weinheim an der Bergstrasse, Germany)

Auteurs

Membres identifiés du Cancéropôle Est :
Dr VILLA Pascal , Dr BONNET Dominique , Dr KLYMCHENKO Andrey

Tous les auteurs :
Florès O, Berthomé Y, Weiss L, Griesbaum-Dubourg S, Riché S, Wagner P, Valencia C, Villa P, Klymchenko AS, Karpenko J, Bonnet D

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/40273184

Résumé

Fluorogenic dimers enable background-free imaging of biological targets under wash-free conditions owing to a strong fluorescence enhancement in the apolar cell microenvironment. However, it is crucial that the imaging probe interacts solely with the target receptor to avoid non-specific interactions and ensure detection with a high signal-to-noise ratio. Herein, we describe a convenient and rapid approach for the synthesis of various functionalized cyanine dyes by click chemistry allowing the fine-tuning of the physicochemical and fluorogenic properties of the dimers. A structure-interaction relationship study was conducted for the fluorogenic dimers in the presence of bovine serum albumin (BSA) and liposomes as models of serum proteins and cell membranes. We identified d-Cy-E which combined the lowest non-specific interactions with the optimal fluorescence turn-on properties. By conjugating d-Cy-E to a peptide ligand of the apelin GPCR, we developed Ap-d-Cy-E, the first fluorescent turn-on probe for the background-free imaging of this receptor in living cells.