Binding of estrogenic compounds to recombinant estrogen receptor-alpha: application to environmental analysis.
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-alphaligand-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-alpha 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-alpha to separate ligands for ER and AhR that are present in river sediments. Immobilized ER-alpha, which does not retain dioxin-like compounds, enabled us to isolate and concentrate ER ligands to facilitate their further analysis.
Pillon
Arnaud
A
INSERM (Institut National de la Santé et de la Recherche Médicale), Unité 540, Montpellier, France.
Boussioux
Anne-Marie
AM
Escande
Aurélie
A
Aït-Aïssa
Sélim
S
Gomez
Elena
E
Fenet
Hélène
H
Ruff
Marc
M
Moras
Dino
D
Vignon
Françoise
F
Duchesne
Marie-Josèphe
MJ
Casellas
Claude
C
Nicolas
Jean-Claude
JC
Balaguer
Patrick
P
D001681:Biological Assay / Q000379:methods; D001943:Breast Neoplasms / Q000473:pathology; D004784:Environmental Monitoring / Q000379:methods*; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D004967:Estrogens / Q000032:analysis* / Q000494:pharmacology*; D005260:Female; D006801:Humans; D008024:Ligands; D010957:Plasmids; D014162:Transfection; D014407:Tumor Cells, Cultured; D014874:Water Pollutants, Chemical / Q000032:analysis* / Q000494:pharmacology*
D047628:Estrogen Receptor alpha; D004967:Estrogens; D008024:Ligands; D014874:Water Pollutants, Chemical
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