Multifaceted Modeling of Estrogen Receptor

雌激素受体的多方面建模

• 批准号: 9892624
• 负责人: Sichun Yang
• 金额: $ 3.04万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9892624
• 关键词: Affect; Biochemical; Biological Assay; Biophysics; Cells; Chemicals; Complex; Crystallization; DNA; DNA Binding Domain; Data; Data Analyses; Disease; Estrogen Receptor alpha; Estrogen Receptors; Family; Genes; Genetic Transcription; Goals; Homeostasis; Hydrophobicity; Individual; Knowledge; Ligand Binding Domain; Ligands; Malignant neoplasm of lung; Measurement; Measures; Mediating; Methodology; Methods; Modeling; Molecular; Molecular Conformation; Mutate; Mutation; Nuclear Hormone Receptors; Outcome; Point Mutation; Protein Region; Proteins; Reporting; Research Personnel; Resolution; Roentgen Rays; Rotation; Sampling; Shapes; Signal Transduction; Site; Site-Directed Mutagenesis; Steroids; Structural Protein; Structure; Sushi Domain; Synchrotrons; System; Tacrolimus Binding Protein 1A; Testing; Transcriptional Regulation; Transforming Growth Factor beta; Uterine Cancer; base; cancer type; computer studies; crosslink; experimental study; flexibility; malignant breast neoplasm; member; mutant; novel therapeutic intervention; protein complex; protein protein interaction; protein structure; receptor; receptor binding; simulation; time use

PROJECT SUMMARY/ABSTRACT The estrogen receptor alpha (ER) is a member of a large nuclear hormone receptor family that regulates a network of genes to control cell homeostasis. Aberrant expression or point mutations of ER have been associated with several types of cancers, but a mechanistic understanding of ER function at the molecular level is still missing. Such knowledge will help to suggest novel therapeutic interventions. The ER mediates steroid control of transcription through the two domains: the DNA-binding domain (DBD) and the ligand-binding domain (LBD). The goal of our project is to eventually understand the allosteric cross-talk between these domains. Compelling evidence from several well-studied NRs indicates that interactions are formed between the two domains. However, it remains completely unknown how the two ER domains communicate with each other and identify protein regions that are involved in allosteric signaling. The ER is a homodimer with multiple domains, and structural flexibility, making it a challenging protein for high-resolution structural studies. To date, high-resolution structures of individual domains, but not of the ER complex containing both domains, have been determined. To understand how the DBD and the LBD interact with each other, we have been developing a multifaceted modeling method that combines information from small-angle X-ray scattering (SAXS) and synchrotron footprinting with the high-resolution structures of the individual domains. We have extensively tested and validated this methodology on a variety of systems with known protein complexes. We demonstrate that the combination of these complementary data yields derived complex structures that are close to the known crystal structures. Using these approaches, we have made significant progress in obtaining structural information for the estrogen receptor (ER) from experimental SAXS and footprinting measurements, and chemical cross-linking. These data will reveal information about the ER that has been unattainable through other methods and will identify interactions between the DBD and the LBD as well as previously uncharacterized DBD-LBD interfaces. We will also attempt to explain how several disease-associated ER mutations influence transcriptional potency of ER, and will provide a molecular characterization of the mutant ER complexes.

项目总结/摘要 雌激素受体α（ER）是一个大的核激素受体家族的成员， 调节基因网络以控制细胞内稳态。异常的表情或点 ER的突变与几种类型的癌症有关，但机制 在分子水平上对ER功能的理解仍然缺失。这些知识将有助于 提示新的治疗干预。ER介导类固醇对转录的控制 通过两个结构域：DNA结合结构域（DBD）和配体结合结构域（LBD）。 我们项目的目标是最终了解这些之间的变构串扰 域.来自几个研究充分的NR的令人信服的证据表明， 在两个领域之间形成。然而，目前还完全不清楚这两个ER 结构域相互通信，并识别参与变构的蛋白质区域， 发信号。 ER是具有多个结构域和结构灵活性的同源二聚体，使其具有挑战性。 蛋白质进行高分辨率结构研究。到目前为止，个人的高分辨率结构 结构域，但不是含有两个结构域的ER复合物，已经确定。到 了解DBD和LBD如何相互作用，我们一直在开发一个 结合小角度X射线散射信息的多面建模方法 （SAXS）和同步加速器足迹与个人的高分辨率结构 域.我们已经在各种系统上广泛地测试和验证了这种方法 与已知的蛋白质复合物。我们证明，这些互补的组合 数据产生了与已知晶体结构接近的衍生复杂结构。使用 这些方法，我们已经取得了重大进展，在获得结构信息的 雌激素受体（ER）从实验SAXS和足迹测量，和化学 交联。这些数据将揭示有关ER的信息， 通过其他方法，并将确定DBD和LBD之间的相互作用，以及 以前未表征的DBD-LBD接口。我们还将试图解释几个 疾病相关的ER突变影响ER的转录能力，并将提供一个 突变ER复合物的分子表征。