MOLECULAR PROBES FOR STEROID RECEPTORS

描述(申请人提供)：这个项目的目的是(1)了解雌激素的各种药理作用的分子基础，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-辅活化子相互作用发挥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-辅调节器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，以及它们与辅助调节蛋白的相互作用，以及这些作用是如何被各种ER配体调节的，包括我们开发的ER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相互作用在这些相互作用中的作用，在招募辅助激活因子和辅助抑制因子方面的竞争，SERM的激动性，如他莫昔芬，以及ERAlpha.ERbeta异源二聚体功能的配体调节。利用基于结构和从头设计的方法，我们将开发小分子共激活物结合抑制剂，它们应该在与抗雌激素不同的水平上阻断雌激素的作用，并可能克服乳腺癌的抗雌激素耐药。核激素受体或辅助调节因子的蛋白质微阵列将以快速、高通量、全基因组的方式分析受体-辅助调节因子相互作用的配体调节。该项目将有助于更好地了解雌激素作用的分子基础，调节雌激素活性的新药物，以及发现新的雌激素和其他核受体配体的有力工具。

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.