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Red Fluorescent Turn-On Ligands for Imaging and Quantifying G Protein-Coupled Receptors in Living Cells

Authors

  • Iuliia A. Karpenko,

    1. Laboratoire d'Innovation Thérapeutique UMR 7200 UdS CNRS, LabEx Medalis, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex (France)
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  • Rémy Kreder,

    1. Laboratoire de Biophotonique et Pharmacologie UMR 7213 UdS CNRS, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex (France)
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  • Christel Valencia,

    1. PCBIS, FMTS, UMS 3286, CNRS-UdS, LabEx Medalis, ESBS Pôle API, Bld Sébastien Brant, 67401 Illkirch Cedex (France)
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  • Dr. Pascal Villa,

    1. PCBIS, FMTS, UMS 3286, CNRS-UdS, LabEx Medalis, ESBS Pôle API, Bld Sébastien Brant, 67401 Illkirch Cedex (France)
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  • Dr. Christiane Mendre,

    1. Institut de Génomique Fonctionnelle, CNRS UMR 5203, INSERM U661, Université de Montpellier I et II, 141 rue de la Cardonille, 34094 Montpellier Cedex 5 (France)
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  • Dr. Bernard Mouillac,

    1. Institut de Génomique Fonctionnelle, CNRS UMR 5203, INSERM U661, Université de Montpellier I et II, 141 rue de la Cardonille, 34094 Montpellier Cedex 5 (France)
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  • Prof. Dr. Yves Mély,

    1. Laboratoire de Biophotonique et Pharmacologie UMR 7213 UdS CNRS, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex (France)
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  • Prof. Dr. Marcel Hibert,

    1. Laboratoire d'Innovation Thérapeutique UMR 7200 UdS CNRS, LabEx Medalis, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex (France)
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  • Dr. Dominique Bonnet,

    Corresponding author
    1. Laboratoire d'Innovation Thérapeutique UMR 7200 UdS CNRS, LabEx Medalis, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex (France)
    • Dominique Bonnet, Laboratoire d'Innovation Thérapeutique UMR 7200 UdS CNRS, LabEx Medalis, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex (France)

      Andrey S. Klymchenko, Laboratoire de Biophotonique et Pharmacologie UMR 7213 UdS CNRS, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex (France)

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  • Dr. Andrey S. Klymchenko

    Corresponding author
    1. Laboratoire de Biophotonique et Pharmacologie UMR 7213 UdS CNRS, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex (France)
    • Dominique Bonnet, Laboratoire d'Innovation Thérapeutique UMR 7200 UdS CNRS, LabEx Medalis, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex (France)

      Andrey S. Klymchenko, Laboratoire de Biophotonique et Pharmacologie UMR 7213 UdS CNRS, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex (France)

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Abstract

Classical fluorescence-based approaches to monitor ligand–protein interactions are generally hampered by the background signal of unbound ligand, which must be removed by tedious washing steps. To overcome this major limitation, we report here the first red fluorescent turn-on probes for a G protein-coupled receptor (oxytocin receptor) at the surface of living cells. The peptide ligand carbetocin was conjugated to one of the best solvatochromic (fluorogenic) dyes, Nile Red, which turns on emission when reaching the hydrophobic environment of the receptor. We showed that the incorporation of hydrophilic octa(ethylene glycol) linker between the pharmacophore and the dye minimized nonspecific interaction of the probe with serum proteins and lipid membranes, thus ensuring receptor-specific turn-on response. The new ligand was successfully applied for background-free imaging and quantification of oxytocin receptors in living cells.

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