title: A multiplexed screening method for agonists and antagonists of the estrogen receptor protein.
abstract: The estrogen receptor (ER) is regarded as a significant drug target because of its important physical and pathological function. In this article, we describe a novel screening method to obtain agonists and antagonists of ER. ER was immobilized onto an aldehyde-modified glass slide. The affinity of Cy3-labeled estradiol for ER protein microarrays was then determined. Two libraries, one containing 29 synthetic compounds and the other with 384 natural products that served as a model, were screened to find new ligands for ER. The IC(50) values obtained for tamoxifen and raloxifene were consistent with those found in the literature (4.85 × 10(-7) M versus 1.74~4.23 × 10(-7) M and 7.58 × 10(-8) M versus 0.89~5.84 × 10(-8) M, respectively). Finally, 65 active ligands (5 synthetic compounds and 60 natural products) of ER were identified. This novel method gave identical results to a conventional fluorescence polarization assay, thus verifying the accuracy of this simultaneous multireceptor screening method based on protein microarrays. The presented method is sensitive, accurate, and reliable, and shows great potential for use in high-throughput drug-screening research.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, China.
authors.forename: Zhonghui
authors.initials: Z
authors.lastname: Li
authors.forename: Ming
authors.initials: M
authors.lastname: Yan
authors.forename: Zhoumin
authors.initials: Z
authors.lastname: Li
authors.forename: Maika
authors.initials: M
authors.lastname: Vuki
authors.forename: Dan
authors.initials: D
authors.lastname: Wu
authors.forename: Fei
authors.initials: F
authors.lastname: Liu
authors.forename: Wenying
authors.initials: W
authors.lastname: Zhong
authors.forename: Luyong
authors.initials: L
authors.lastname: Zhang
authors.forename: Danke
authors.initials: D
authors.lastname: Xu
mesh_terms: D045166:Estradiol Congeners / Q000737:chemistry / Q000378:metabolism; D004965:Estrogen Antagonists / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D020849:Raloxifene Hydrochloride / Q000737:chemistry / Q000378:metabolism; D011960:Receptors, Estrogen / Q000819:agonists* / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D013629:Tamoxifen / Q000737:chemistry / Q000378:metabolism
chemical_list: D045166:Estradiol Congeners; D004965:Estrogen Antagonists; D008024:Ligands; D011960:Receptors, Estrogen; D013629:Tamoxifen; D020849:Raloxifene Hydrochloride