Click‐Functionalized Cyanine Fluorogenic Dimers for Improved Detection of GPCRs: Application to Imaging of ApelinR in Living Cells
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 nonspecific 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 nonspecific 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.
Authors
- Stéphanie Riché (ORCID: https://orcid.org/0000-0003-2783-0293)
- Pascal Villa (ORCID: https://orcid.org/0000-0003-3466-4852)
- Julie Karpenko (ORCID: https://orcid.org/0000-0002-7633-3691)
- Dominique Bonnet (ORCID: https://orcid.org/0000-0002-8252-9199)
- Océane Florès (ORCID: https://orcid.org/0000-0003-0156-728X)
- Andrey S. Klymchenko (ORCID: https://orcid.org/0000-0002-2423-830X)
- Yann Berthomé
- Patrick Wagner (ORCID: https://orcid.org/0000-0002-8273-8329)
- C. Alexander Valencia (ORCID: https://orcid.org/0000-0002-5249-6779)
- Lucille Weiss
- Sarah Griesbaum-Dubourg
Institutions
- Centre National de la Recherche Scientifique (FR)
- Centre National pour la Recherche Scientifique et Technique (CNRST) (MA)
- Laboratoire de Biophotonique et Pharmacologie (FR)
- Institut de Chimie de Strasbourg (FR)
- Laboratoire d'Innovation Thérapeutique (FR)
- Plateforme de chimie biologique intégrative de Strasbourg
- Université de Strasbourg (FR)
Publication Details
- Journal
- Chemistry - A European Journal
- Published
- 2025-04-24
- DOI
- https://doi.org/10.1002/chem.202500379
- Citations
- 1
- Primary Topic
- Apelin-related biomedical research
- Type
- article
- Field-Weighted Citation Impact
- 1.39
Funders
- Agence Nationale de la Recherche
- Institut National de la Santé et de la Recherche Médicale
- Université de Strasbourg
- Centre National de la Recherche Scientifique
- Société d'Accélération du Transfert de Technologies