Binding of Estrogenic Compounds to Recombinant Estrogen Receptor-: Application to Environmental Analysis Arnaud Pillon,1 Anne-Marie Boussioux,1 Aurélie Escande,1 Sélim Aït-Aïssa,2 Elena Gomez,3 Hélène Fenet,3 Marc Ruff,4 Dino Moras,4 Françoise Vignon,1 Marie-Josèphe Duchesne,1 Claude Casellas,3 Jean-Claude Nicolas,1 and Patrick Balaguer1 1INSERM (Institut National de la Santé et de la Recherche Médicale), Unité 540, Montpellier, France; 2INERIS (Institut National de l'Environnement Industriels et des Risques), Unité Evaluation des Risques Ecotoxicologiques, Verneuil-en-Halatte, France; 3 UMR 5569 "Hydrosciences," Département des Sciences de l'Environnement et Santé Publique, Faculté de Pharmacie, Montpellier, France; 4IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire), Laboratoire de Biologie et Génomique Structurale, Illkirch, France Abstract Estrogenic activity in environmental samples could be mediated through a wide variety of compounds and by various mechanisms. High-affinity compounds for estrogen receptors (ERs) , such as natural or synthetic estrogens, as well as low-affinity compounds such as alkylphenols, phthalates, and polychlorinated biphenyls are present in water and sediment samples. Furthermore, compounds such as polycyclic aromatic hydrocarbons, which do not bind ERs, modulate estrogen activity by means of the aryl hydrocarbon receptor (AhR) . In order to characterize compounds that mediate estrogenic activity in river water and sediment samples, we developed a tool based on the ER-ligand-binding domain, which permitted us to estimate contaminating estrogenic compound affinities. We designed a simple transactivation assay in which compounds of high affinity were captured by limited amounts of recombinant ER- and whose capture led to a selective inhibition of transactivation. This approach allowed us to bring to light that water samples contain estrogenic compounds that display a high affinity for ERs but are present at low concentrations. In sediment samples, on the contrary, we showed that estrogenic compounds possess a low affinity and are present at high concentration. Finally, we used immobilized recombinant ER- to separate ligands for ER and AhR that are present in river sediments. Immobilized ER-, which does not retain dioxin-like compounds, enabled us to isolate and concentrate ER ligands to facilitate their further analysis. Key words: aryl hydrocarbon receptor, bioluminescent cell lines, environmental samples, estrogen receptor, xenoestrogens. Environ Health Perspect 113: 278-284 (2005) . doi:10.1289/ehp.7522 available via http://dx.doi.org/ [Online 9 December 2004] Address correspondence to P. Balaguer, INSERM, Unité 540, 60 rue de Navacelles, 34090 Montpellier, France. Telephone: 33-467043703. Fax: 33-467540598. E-mail: balaguer@montp.inserm.fr A.P. is financially supported by a grant from the Languedoc-Roussillon Région and the Institut National de l'Environnement Industriels et des Risques. The authors declare they have no competing financial interests. Received 25 August 2004 ; accepted 9 December 2004. The full version of this article is available for free in HTML or PDF formats. |