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Nile Red-Based GPCR Ligands as Ultrasensitive Probes of the Local Lipid Microenvironment of the Receptor.pdf (1.16 MB)

Nile Red-Based GPCR Ligands as Ultrasensitive Probes of the Local Lipid Microenvironment of the Receptor

preprint
submitted on 17.11.2020, 12:37 and posted on 18.11.2020, 04:22 by Fabien Hanser, Claire Marsol, Christel Valencia, Pascal Villa, Andrey S. Klymchenko, Dominique Bonnet, Iuliia Karpenko

The local lipid microenvironment of transmembrane receptors is an essential factor in 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 non-peptidic 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 non-specific 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 the information about the local polarity and the lipid order via the wavelength-shifting emission of the Nile Red fluorophore.

Funding

LabEx MEDALIS (ANR-10-LABX-0034)

Pierre Fabre and LabEx MEDALIS Attractivity Grant

CNRS France

University of Strasbourg, France

History

Email Address of Submitting Author

i.karpenko@unistra.fr

Institution

Laboratoire d’Innovation Thérapeutique, LabEx MEDALIS, UMR7200, CNRS/Université de Strasbourg

Country

France

ORCID For Submitting Author

0000-0002-7633-3691

Declaration of Conflict of Interest

No conflict of interest

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