title: Monitoring ligand-induced conformational changes for the identification of estrogen receptor agonists and antagonists.
abstract: Nuclear receptors are transcription factors that are important targets for current drug discovery efforts as they play a role in many pathological processes. Their activity can be regulated by small molecules like hormones and drugs that can have agonistic or antagonistic functions. These ligands bind to the receptor and account for diverse conformational changes that are crucial determinants for the receptor activity. Here, we set out to develop FLiN (fluorescent labels in nuclear receptors), a direct binding assay that detects conformational changes in the estrogen receptor. The assay is based on the introduction of a cysteine residue and subsequent specific labeling of the receptor with a thiol-reactive fluorophore. Changes in the receptor conformation upon ligand binding lead to differences in the microenvironment of the fluorophore and alter its emission spectrum. The FLiN assay distinguishes between different binding modes and is suitable for high-throughput screening.
authors: Technische Universität Dortmund, Fakultät für Chemie und Chemische Biologie, Otto-Hahn-Strasse 6, 44227 Dortmund (Germany).
authors: Svenja C
authors: SC
authors: Mayer-Wrangowski
authors: Daniel
authors: D
authors: Rauh
mesh_terms: D065171:Estrogen Receptor Antagonists / Q000737:chemistry*; D004967:Estrogens / Q000737:chemistry*; D008024:Ligands; D008968:Molecular Conformation
chemical_list: D065171:Estrogen Receptor Antagonists; D004967:Estrogens; D008024:Ligands
keywords: direct binding assay; drug discovery; estrogen receptor; fluorescence spectroscopy