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Stimulation of human olfactory receptor 17-40 with odorants probed by surface plasmon resonance

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Abstract

We report here the results of human olfactory receptor (OR) 17-40 stimulation with some odorants probed by means of the double-channel surface plasmon resonance platform NanoSPR-6. OR 17-40 tagged with N-terminal cmyc sequence was heterologously co-expressed with Gαolf protein in yeast, and receptor-carrying nanosomes were prepared from yeast membrane fraction. Then, receptors were specifically captured via anti-cmyc antibody attached to the gold-coated substrate in orientated or random way. Measurement of odorants effects were carried out in the presence of GTP-γ-S in differential mode in order to compensate bulk changes of refractive index. For the first time, biosensing efficiency of olfactory films was discussed in terms of their thickness and Gαolf accessibility to GTP-γ-S. Bell-shaped response profile with two maxima (near 1 nM and near 1 μM) was established for helional, which is documented as highly specific agonist of OR 17-40. Unrelated odorant heptanal used as control, did not evoke significant variations of differential signal.

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Abbreviations

GPCRs:

G protein-coupled receptors

ORs:

Olfactory receptors

SAM:

Self-assembled monolayer

SPR:

Surface plasmon resonance

PBS:

Phosphate-buffered saline

AFM:

Atomic force microscopy

CV:

Cyclic voltammetry

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Acknowledgments

Vladimir I. Chegel appreciates financial support from NATO CLG grant PDD (CP)-(CBP.NUKR.CLG981776). Irina Benilova was supported by EGIDE (France) through the doctoral Bourse Eiffel.

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Authors and Affiliations

  1. Laboratoire Ampère, Ecole Centrale de Lyon, 36 Avenue Guy de Collongue, 69134, Ecully Cedex, France

    Irina Benilova & Claude Martelet

  2. Lashkaryov Institute of Semiconductor Physics NASU, 45 Avenue Nauki, 03028, Kyiv, Ukraine

    Vladimir I. Chegel & Yuri V. Ushenin

  3. Neurobiologie de l’Olfaction et de la Prise Alimentaire, Récepteurs et Communication Chimique, INRA, Domaine de Vilvert, 78352, Jouy-en-Josas Cedex, France

    Jasmina Vidic & Edith Pajot

  4. Laboratory of Biomolecular Electronics, Institute of Molecular Biology and Genetics NASU, 150 Zabolotnogo Street, 03143, Kyiv, Ukraine

    Irina Benilova & Alexey P. Soldatkin

  5. Laboratoire des Sciences Analytiques, UMR-CNRS 5180, Université Claude Bernard, Lyon 1, 43 Boulevard du 11 Novembre 1918, 69622, Villeurbanne Cedex, France

    Nicole Jaffrezic-Renault

Authors
  1. Irina Benilova
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  2. Vladimir I. Chegel
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  3. Yuri V. Ushenin
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  4. Jasmina Vidic
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  6. Claude Martelet
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  7. Edith Pajot
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  8. Nicole Jaffrezic-Renault
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Correspondence to Irina Benilova.

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Benilova, I., Chegel, V.I., Ushenin, Y.V. et al. Stimulation of human olfactory receptor 17-40 with odorants probed by surface plasmon resonance. Eur Biophys J 37, 807–814 (2008). https://doi.org/10.1007/s00249-008-0272-5

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  • DOI: https://doi.org/10.1007/s00249-008-0272-5

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