Nile Red-Based GPCR Ligands as Ultrasensitive Probes of the Local Lipid Microenvironment of the ReceptorClick to copy article linkArticle link copied!
- Fabien HanserFabien HanserLaboratoire d’Innovation Thérapeutique, UMR7200 CNRS/Université de Strasbourg, Strasbourg Drug Discovery and Development Institute (IMS), 74 route du Rhin, 67401 Illkirch-Graffenstaden, FranceMore by Fabien Hanser
- Claire MarsolClaire MarsolLaboratoire d’Innovation Thérapeutique, UMR7200 CNRS/Université de Strasbourg, Strasbourg Drug Discovery and Development Institute (IMS), 74 route du Rhin, 67401 Illkirch-Graffenstaden, FrancePlate-forme de chimie biologique intégrative de Strasbourg (PCBiS), UMS 3286 CNRS/Université de Strasbourg, Strasbourg Drug Discovery and Development Institute (IMS), ESBS Pôle API, Bld Sébastien Brant, 67412 Illkirch-Graffenstaden, FranceMore by Claire Marsol
- Christel ValenciaChristel ValenciaPlate-forme de chimie biologique intégrative de Strasbourg (PCBiS), UMS 3286 CNRS/Université de Strasbourg, Strasbourg Drug Discovery and Development Institute (IMS), ESBS Pôle API, Bld Sébastien Brant, 67412 Illkirch-Graffenstaden, FranceMore by Christel Valencia
- Pascal VillaPascal VillaPlate-forme de chimie biologique intégrative de Strasbourg (PCBiS), UMS 3286 CNRS/Université de Strasbourg, Strasbourg Drug Discovery and Development Institute (IMS), ESBS Pôle API, Bld Sébastien Brant, 67412 Illkirch-Graffenstaden, FranceMore by Pascal Villa
- Andrey S. KlymchenkoAndrey S. KlymchenkoLaboratoire de Bioimagerie et Pathologies, UMR 7021 CNRS/Université de Strasbourg, 74 route du Rhin, 67401 Illkirch-Graffenstaden, FranceMore by Andrey S. Klymchenko
- Dominique Bonnet*Dominique Bonnet*E-mail: [email protected]Laboratoire d’Innovation Thérapeutique, UMR7200 CNRS/Université de Strasbourg, Strasbourg Drug Discovery and Development Institute (IMS), 74 route du Rhin, 67401 Illkirch-Graffenstaden, FranceMore by Dominique Bonnet
- Julie Karpenko*Julie Karpenko*E-mail: [email protected]Laboratoire d’Innovation Thérapeutique, UMR7200 CNRS/Université de Strasbourg, Strasbourg Drug Discovery and Development Institute (IMS), 74 route du Rhin, 67401 Illkirch-Graffenstaden, FranceMore by Julie Karpenko
Abstract

The local lipid microenvironment of transmembrane receptors is an essential factor in G protein coupled receptor (GPCR) signaling. However, tools are currently missing for studying endogenously expressed GPCRs in primary cells and tissues. Here, we introduce fluorescent environment-sensitive GPCR ligands for probing the microenvironment of the receptor in living cells using fluorescence microscopy under no-wash conditions. We designed and synthesized antagonist ligands of the oxytocin receptor (OTR) by conjugating a high-affinity nonpeptidic OTR ligand PF-3274167 to the environment-sensitive fluorescent dye Nile Red. The length of the polar PEG spacer between the pharmacophore and the fluorophore was adjusted to lower the nonspecific interactions of the probe while preserving a strong fluorogenic response. We demonstrated that the new probes embed into the lipid bilayer in the vicinity of the receptor and convey information about the local polarity and the lipid order via the wavelength-shifting emission of the Nile Red fluorophore.
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