Han, Ki-Cheol Kim, Jung Hwan Kim, Kook-Han Kim, Eunice EunKyeong Seo, Jin-Ho Yang, Eun Gyeong 2024-01-20T19:33:58Z 2024-01-20T19:33:58Z 2021-09-02 2010-03-15 0003-2697 https://pubs.kist.re.kr/handle/201004/131630 Farnesoid X receptor (FXR) serves as a receptor for chenodeoxycholic acid (CDCA) and other bile acids, and it coordinates cholesterol and lipid metabolism. Because targeting the FXR-CDCA interaction might provide a way to regulate lipid homeostasis, we developed an FXR binding assay based on fluorescence polarization. Employing a fluorescently labeled CDCA (CDCA-F), we showed that CDCA-F selectively bound to the ligand binding domain of FXR (FXR-LBD) among nuclear receptors. The assay was then used for screening inhibitors against the FXR-CDCA interaction, thereby discovering four relatively potent inhibitors. The selected inhibitors were further studied for changes in intrinsic tryptophan fluorescence of FXR-LBD to gain structural insights into the interaction. Furthermore, transactivation effects of the inhibitors on the human bile salt excretory pump (BSEP) promoter were examined to reveal their cellular activities in the FXR-mediated pathway. Therefore, we demonstrated that the developed assay would offer an efficient primary screening tool for identifying FXR modulators. (C) 2010 Elsevier Inc. All rights reserved. English ACADEMIC PRESS INC ELSEVIER SCIENCE SALT EXPORT PUMP ACTIVATED RECEPTOR BILE-ACIDS NUCLEAR RECEPTOR COACTIVATOR ASSOCIATION PROTEIN GENE FATTY-ACIDS LIGANDS BINDING HEXACHLOROPHENE Identification of farnesoid X receptor modulators by a fluorescence polarization-based interaction assay Article 10.1016/j.ab.2009.11.008 1 ANALYTICAL BIOCHEMISTRY, v.398, no.2, pp.185 - 190 ANALYTICAL BIOCHEMISTRY 398 2 185 190 scie scopus 000274615100006 2-s2.0-77649192410 Biochemical Research Methods Biochemistry & Molecular Biology Chemistry, Analytical Biochemistry & Molecular Biology Chemistry Article SALT EXPORT PUMP ACTIVATED RECEPTOR BILE-ACIDS NUCLEAR RECEPTOR COACTIVATOR ASSOCIATION PROTEIN GENE FATTY-ACIDS LIGANDS BINDING HEXACHLOROPHENE Farnesoid X receptor Chenodeoxycholic acid Fluorescence polarization Antagonist Agonist Reporter assay