MOLECULAR PROBES FOR STEROID RECEPTORS

描述（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（SERM）和抗雌激素，（2）开发通过阻断受体-辅激活剂相互作用发挥作用的新型雌激素作用抑制剂，以及（3）制备蛋白质微阵列，其可用于研究配体如何在全基因组基础上调节核受体-辅调节因子相互作用。我们将应用先进的荧光方法来研究雌激素受体ER α和ER β的构象动力学和相互作用，以及它们与辅助调节蛋白的相互作用，以及这些如何被各种ER配体（包括我们开发的ER亚型选择性配体）所调节。具体的研究将集中在蛋白质构象动力学和互惠在这些相互作用中的作用，竞争招募的coactivators与corepressors，激动剂的SERMs，如他莫昔芬，和配体调节ER α的功能。ER β异二聚体。使用基于结构的和从头设计，我们将开发小分子辅激活因子结合抑制剂，这些抑制剂应该在与抗雌激素不同的水平上阻断雌激素作用，并且可能克服乳腺癌中的抗雌激素耐药性。将开发核激素受体或辅助调节因子的蛋白质微阵列，以快速、高通量、全基因组方式测定受体-辅助调节因子相互作用的配体调节。该项目将导致对雌激素作用的分子基础的更好理解，调节雌激素活性的新型药物，以及发现新型雌激素和其他核受体配体的强大工具。

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.