title: Estrogen receptor affinity chromatography: a new method for characterization of novel estrogenic disinfection by-products.
abstract: To identify the unknown estrogenic disinfection by-products (DBPs) from the chlorination extract, an effective method based on affinity chromatography with immobilized human recombinant estrogen receptor α (ERα) was developed, which has an advantage in targeting different potential estrogenic compounds from mixed sample simultaneously by comparing their relative binding activities to ER. The new method worked well for six known environmental estrogens. To further test the validity of this method for unknown chemicals, six DBPs of diethylstilbestrol (DES) with relatively strong ER binding affinity after chlorination were isolated and identified. It was found that except for 2-chloro-DES which showed 1.36 times stronger binding affinity than DES, most of the by-products bound to ER much more weakly than DES. All these seven by-products induced a dose-dependent transcriptional activation in two-hybrid-yeast assays. Z,Z-dienestrol (DE) and 2-chloro-DES, which exhibiting the weakest and the strongest binding affinity, were further tested for their transcriptional potential as 0.00243 and 0.014 compared to DES, respectively. However, they were still potential harmful environmental estrogenic disruptors as their estrogenic activities were much stronger than that of bisphenol A (BPA). These results demonstrated that the new method can help to screen unknown estrogenic compounds from mixture more efficiently.
authors.affiliation: Environmental Simulation and Pollution Control State Key Joint Laboratory, School of Environment, Tsinghua University, Beijing 100084, China.
authors.forename: Guodong
authors.initials: G
authors.lastname: Shang
authors.affiliation: Environmental Simulation and Pollution Control State Key Joint Laboratory, School of Environment, Tsinghua University, Beijing 100084, China.
authors.forename: Jinling
authors.initials: J
authors.lastname: Xue
authors.affiliation: Environmental Simulation and Pollution Control State Key Joint Laboratory, School of Environment, Tsinghua University, Beijing 100084, China.
authors.forename: Man
authors.initials: M
authors.lastname: Li
authors.affiliation: Environmental Simulation and Pollution Control State Key Joint Laboratory, School of Environment, Tsinghua University, Beijing 100084, China; Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China.
authors.forename: Hong-Ying
authors.initials: HY
authors.lastname: Hu
authors.affiliation: Environmental Simulation and Pollution Control State Key Joint Laboratory, School of Environment, Tsinghua University, Beijing 100084, China; Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China. Electronic address: luyun@tsinghua.edu.cn.
authors.forename: Yun
authors.initials: Y
authors.lastname: Lu
mesh_terms: D001559:Benzhydryl Compounds / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D004054:Diethylstilbestrol / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D004203:Disinfection / Q000379:methods; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D004967:Estrogens / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D054879:Halogenation; D006801:Humans; D055767:Immobilized Proteins / Q000378:metabolism*; D010636:Phenols / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D000098826:Bisphenol A Compounds
chemical_list: D001559:Benzhydryl Compounds; D004054:Diethylstilbestrol; D047628:Estrogen Receptor alpha; D004967:Estrogens; D055767:Immobilized Proteins; D010636:Phenols; D011994:Recombinant Proteins; C506487:ESR1 protein, human; C006780:bisphenol A; D000098826:Bisphenol A Compounds
keywords: Affinity chromatography; Diethylstilbestrol; Disinfection by-product; Estrogen screening; Estrogenicity