Electrochemical detection of xenoestrogenic and antiestrogenic compounds using a yeast two-hybrid-17-beta-estradiol system.
The goal of this study was to determine the effects of various compounds on the 17-beta-estradiol-induced dimerization of the human estrogen receptor alpha (hERalpha), a nuclear transcription factor. For this purpose, we used a modified yeast two-hybrid (YTH) bioassay designed to study protein-protein interactions, based on the electrochemical monitoring of hERalpha dimerization and detected as beta-D-galactosidase reporter gene activity in a synthetic substrate p-aminophenyl-beta-D-galactopyranoside (pAPG). Compared with 17-beta-estradiol activity, genistein, bisphenol-A (BPA), and naringenin induced dimerization to a lower extent by four, five and six magnitudes of orders of magnitude, respectively. In the presence of physiological concentrations of 17-beta-estradiol, both tamoxifen and the analgesic drug acetaminophen inhibited hER dimerization in an antiestrogenic manner.
Schwartz-Mittelman
Adrian
A
Department of Molecular Microbiology and Biotechnology, Tel-Aviv University, Ramat-Aviv 69978, Israel.
Baruch
Anat
A
Neufeld
Tova
T
Buchner
Virginia
V
Rishpon
Judith
J
D000082:Acetaminophen / Q000494:pharmacology; D001559:Benzhydryl Compounds; D019281:Dimerization; D004563:Electrochemistry / Q000379:methods*; D004958:Estradiol / Q000494:pharmacology*; D020847:Estrogen Receptor Modulators / Q000494:pharmacology*; D047628:Estrogen Receptor alpha / Q000737:chemistry / Q000187:drug effects*; D044950:Flavanones / Q000494:pharmacology; D017930:Genes, Reporter; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D010636:Phenols / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D020798:Two-Hybrid System Techniques; D000098826:Bisphenol A Compounds
D000082:Acetaminophen; D001559:Benzhydryl Compounds; D004958:Estradiol; D020847:Estrogen Receptor Modulators; D047628:Estrogen Receptor alpha; D044950:Flavanones; D019833:Genistein; D010636:Phenols; C005273:naringenin; C006780:bisphenol A; D000098826:Bisphenol A Compounds