MOLECULAR PROBES FOR STEROID RECEPTORS

描述（由申请人提供）：该项目的目的是(1)了解雌激素的不同药理学的分子基础，包括选择性雌激素受体调节剂（SERMs）和抗雌激素，(2)开发通过阻断受体-辅激活剂相互作用的新型雌激素作用抑制剂，以及(3)制备可用于研究配体如何在全基因组基础上调节核受体-辅激活剂相互作用的蛋白质微阵列。我们将应用先进的荧光方法来研究雌激素受体erα和erβ的构象动力学和相互作用，以及它们与协同调节蛋白的相互作用，以及它们如何被各种内质网配体调节，包括我们开发的内质网亚型选择性配体。具体的研究将集中在蛋白质构象动力学和互易性在这些相互作用中的作用，共激活因子与共抑制因子招募的竞争，serm（如他莫昔芬）的激动作用，以及er - α功能的配体调节。ERbeta形成。使用基于结构和从头设计，我们将开发小分子共激活因子结合抑制剂，它应该在不同程度上阻断雌激素的作用，而不是抗雌激素，并可能克服乳腺癌的抗雌激素抵抗。核激素受体或共调节剂的蛋白质微阵列将被开发出来，以快速、高通量、全基因组的方式检测受体-共调节剂相互作用的配体调节。该项目将有助于提高对雌激素作用的分子基础的理解，为发现新的雌激素和核受体的其他配体提供强有力的工具。

DESCRIPTION (provided by applicant): The aims of this project are (1) to understand the molecular basis of the diverse pharmacology of estrogens, including the selective estrogen receptor modulators (SERMs) and antiestrogens, (2) to develop novel inhibitors of estrogen action that act by blocking receptor-coactivator interactions, and (3) to prepare protein microarrays that can be used to study how ligands regulate nuclear receptor-coregulator interactions on a genome-wide basis. We will apply advanced fluorescence methodologies to study the conformational dynamics and interactions of the estrogen receptors ERalpha and ERbeta and their interactions with coregulator proteins and how these are modulated by various ER ligands, including the ER subtype-selective ligands we have developed. Specific studies will focus on the role of protein conformational dynamics and reciprocity in these interactions, competition in recruitment of coactivators vs. corepressors, the agonism of SERMs such as tamoxifen, and ligand regulation of the function of ERalpha.ERbeta heterodimers. Using structure-based and de novo design, we will develop small molecule coactivator binding inhibitors that should block estrogen action at a different level than antiestrogens and might overcome antiestrogen resistance in breast cancer. Protein microarrays of nuclear hormone receptors or coregulators will be developed to assay ligand regulation of receptor-coregulator interaction in a rapid, high throughput, genome-wide manner. This project should lead to improved understanding of the molecular basis of estrogen action, novel agents to regulate estrogen activity, and powerful tools for the discovery of novel estrogens and other ligands for nuclear receptors.