Affinity purification using recombinant PXR as a tool to characterize environmental ligands

Authors

  • Sonia Dagnino,

    Corresponding author
    1. IRCM, Institut de Recherche en Cancerologie de Montpellier, CRLC Val d'Aurelle Paul Lamarque, INSERM, U896 34298 Montpellier, France
    2. Université Montpellier 1, 34298 Montpellier, France
    3. UMR 5569 Hydrosciences, Université Montpellier I, 34060 Montpellier, France
    Current affiliation:
    1. U.S. EPA, Office of Research and Development, National Exposure Research Laboratory, NC 27 711, USA
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  • Virginie Bellet,

    1. IRCM, Institut de Recherche en Cancerologie de Montpellier, CRLC Val d'Aurelle Paul Lamarque, INSERM, U896 34298 Montpellier, France
    2. Université Montpellier 1, 34298 Montpellier, France
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  • Marina Grimaldi,

    1. IRCM, Institut de Recherche en Cancerologie de Montpellier, CRLC Val d'Aurelle Paul Lamarque, INSERM, U896 34298 Montpellier, France
    2. Université Montpellier 1, 34298 Montpellier, France
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  • Anne Riu,

    1. UMR 1089 Xénobiotiques, INRA, F-31931 Toulouse, France
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  • Sélim Aït-Aïssa,

    1. Institut National de l'Environnement Industriels et des Risques (INERIS), Unité Ecotoxicologie in vitro et in vivo f-60550 Verneuil-en-Halatte, France
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  • Vincent Cavaillès,

    1. IRCM, Institut de Recherche en Cancerologie de Montpellier, CRLC Val d'Aurelle Paul Lamarque, INSERM, U896 34298 Montpellier, France
    2. Université Montpellier 1, 34298 Montpellier, France
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  • Hélène Fenet,

    1. UMR 5569 Hydrosciences, Université Montpellier I, 34060 Montpellier, France
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  • Patrick Balaguer

    Corresponding author
    1. IRCM, Institut de Recherche en Cancerologie de Montpellier, CRLC Val d'Aurelle Paul Lamarque, INSERM, U896 34298 Montpellier, France
    2. Université Montpellier 1, 34298 Montpellier, France
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Abstract

Many environmental endocrine disrupting compounds act as ligands for nuclear receptors. The human pregnane X receptor (hPXR), for instance, is activated by a variety of environmental ligands such as steroids, pharmaceutical drugs, pesticides, alkylphenols, polychlorinated biphenyls and polybromo diethylethers. Some of us have previously reported the occurrence of hPXR ligands in environmental samples but failed to identify them. The aim of this study was to test whether a PXR-affinity column, in which recombinant hPXR was immobilized on solid support, could help the purification of these chemicals. Using PXR ligands of different affinity (10 nM < EC50 < 10 μM), we demonstrated that the PXR-affinity preferentially column captured ligands with medium to high affinities (EC50 < 1 μM). Furthermore, by using the PXR-affinity column to analyze an environmental sample containing ERα, AhR, AR, and PXR activities, we show that (i) half of the PXR activity of the sample was due to compounds with medium to high affinity for PXR and (ii) PXR shared ligands with ERα, AR, and AhR. These findings demonstrate that the newly developed PXR-affinity column coupled to reporter cell lines represents a valuable tool for the characterization of the nature of PXR active compounds and should therefore guide and facilitate their further analysis. © 2012 Wiley Periodicals, Inc. Environ Toxicol 29: 207–215, 2014.

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