STRUCTURAL STUDIES OF THE DNA-BINDING AND LIGAND-BINDING DOMAINS OF THE HUMAN

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 自2005年春季以来，我们在APS收集了与ERAlpha配体结合域(LBD)相关的结构数据，这些LBD与新的配体络合，这些配体对ERAlpha和ERbeta的效力和选择性不同。从这些数据中，我们能够推测这些化合物与Erb结合的一种可能模式，在溶液中它们对ERb具有更高的选择性。这些OBCP化合物及其新颖的3D支架正在评估其各种应用，因为它们的生物学效应从完全激动剂、部分激动剂/拮抗剂或完全拮抗剂，取决于组织和靶基因启动子的背景。 我们继续我们的结构-功能表征的两个已知的ER亚型与ERbeta LBD和OBCP化合物的晶体络合物。这些结构将加深我们对如何更好地利用OBCP支架开发改进的亚型选择性激动剂和/或新型拮抗剂的理解。 另一个重要的进展是，我们终于得到了一个ERbeta截断突变体的衍射性晶体，它包括DNA结合域(DBD)、铰链区和随后的LBD。这将是一个高分辨率的结构，因为当我们在上一次运行结束前测试它时，它的衍射率达到了1.34A。这是一个令人兴奋的结构，因为到目前为止，还没有既包括DBD又包括LBD的ERpha/beta结构。确定这种结构将使我们更接近于了解整个ERbeta蛋白的结构-功能调节，ERbeta蛋白是一种具有多个结构域的重要治疗分子。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Since Spring 2005 we collected structural data at the APS which related to ERalpha ligand-binding domain (LBD) complexed with novel ligands that vary in their potency and selectivity for ERalpha and ERbeta. From these data, we were able to speculate on a possible mode of binding of these compounds to ERb for which they are more selective in solution. These OBCP compounds with their novel 3D scaffolds are being evaluated for various applications since their biological effects from being full agonists, partial agonists/antagonists, or complete antagonists, depending on the tissue and target gene promoter context. We continue our structure-function characterization of the two known ER subtypes with crystal complexes of ERbeta LBD and the OBCP compounds. These structures will further our understanding of how to better use the OBCP scaffold for the development of improved subtype selective agonists and/or novel antagonists. Another important development is that we finally have diffraction quality crystals of an ERbeta truncation mutant that includes the DNA-binding domain (DBD), a hinge region and the following LBD. This promises to be a high resolution structure since it diffracted to 1.34A when we tested it just before the end of the previous run. This is an exciting construct because, to date, there are no available structures of ERalpha/beta that include both the DBD and LBD. Determining this structure will bring us closer to understanding the structure-function regulation of the entire ERbeta protein, a therapeutically important molecule with multiple domains.