Mechanistic Studies of Ligand-Regulated Nuclear Receptor RXR Activity

配体调节核受体 RXR 活性的机制研究

基本信息

批准号：
7659605
负责人：
Yong Li
金额：
$ 18.94万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-17 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7659605
关键词：
Address Adverse effects Affinity Amino Acid Sequence Binding Binding Sites Biochemical Biological Assay Biological Process C-terminal Cells Chemicals Clinical Complex Consensus DNA DNA Binding DNA Binding Domain DNA Structure DNA-Protein Interaction Development Disease Drug Delivery Systems Elements Family member Foundations Gene Targeting Goals Homeostasis Human Indium Knowledge Ligand Binding Ligand Binding Domain Ligands Mediating Metabolic Metabolic Diseases Metabolism Modeling Molecular Mutation N-terminal Nuclear Receptors Pathway interactions Peroxisome Proliferator-Activated Receptors Pharmaceutical Preparations Physiological Physiology Proteins RXR Receptor Gene Regulation Relative (related person)Reporting Research Response Elements Retinoids Role Signal Transduction Site Specificity Structure Testing Therapeutic Uses Thyroid Hormones Toxic effect Validation Vitamin A Vitamin D X-Ray Crystallography alitretinoin base cofactor cross reactivity design drug discovery effective therapy human disease improved insight intermolecular interaction multidisciplinary public health relevance receptor receptor binding response three dimensional structure

项目摘要

DESCRIPTION (provided by applicant): The goal of our research is to unravel the molecular basis of the nuclear receptor actions using X-ray crystallography in conjunction with multidisciplinary molecular and biochemical approaches. The nuclear receptor retinoid X receptors (RXRs) are key ligand-activated transcriptional regulators involved in a wide range of human physiology, including development and metabolism. RXRs are well-established drug targets. However, the clinical use of RXR ligands is clearly tempered by side effects like toxicity, possibly associated with their low selectivity and affinity and also the cross-reactivity with other functions of RXRs. To understand the molecular basis of ligand-mediated signaling of RXRs in human physiology, we are planning to determine the crystal structures of a RXR multi-domain fragment bound to a DNA response fragment of their target genes. Following structure determination, we will perform mutational studies, biochemical analysis, and cell-based transcriptional assays to confirm functional significance of key structure elements. This study will reveal the molecular basis of the functional specificity of RXRs and its ligands at key regulatory steps: from ligand binding and to target gene recognition. The structural insights will provide a rational template for structure-based drug discovery of highly selective and efficient RXR ligands with reduced side effects for the therapeutic use in human disease. Moreover, the structure generated in this project will serve as a model to study the ligand-dependent regulation of other nuclear receptor family members. PUBLIC HEALTH RELEVANCE: This proposal addresses a critical need in the current nuclear receptor research by providing three-dimensional structures of the multi-domain retinoid X receptor bound to its target genes. The structural and functional studies will reveal detailed insights on how drug specificity and affinity can be further improved, thus providing a foundation for rational drug discovery for effective treatment of human disease like metabolic disease.

描述（由申请人提供）：我们的研究的目的是使用X射线晶体学结合多学科分子和生化方法来阐明核受体作用的分子基础。核受体类维生素类动物X受体（RXR）是参与广泛人类生理学的主要配体激活的转录调节剂，包括发育和代谢。 RXR是公认的药物靶标。然而，RXR配体的临床使用显然会被毒性（例如毒性），可能与其选择性低和亲和力以及与其他RXR功能的交叉反应性有关。为了了解RXR在人类生理学中的配体介导的信号的分子基础，我们计划确定与其靶基因的DNA响应片段结合的RXR多域片段的晶体结构。遵循结构确定，我们将进行突变研究，生化分析和基于细胞的转录测定，以确认关键结构元素的功能意义。这项研究将揭示RXRS及其配体功能特异性的分子基础：从配体结合和靶向基因识别。结构性见解将为基于结构的药物发现高度选择性和有效的RXR配体提供合理的模板，并降低了在人类疾病中治疗的副作用。此外，该项目中产生的结构将成为研究其他核受体家族成员配体依赖性调节的模型。公共卫生相关性：该提案通过提供与其靶基因结合的多域类维生素类X受体的三维结构来满足当前核受体研究的关键需求。结构和功能研究将揭示有关如何进一步改善药物特异性和亲和力的详细见解，从而为有效治疗人类疾病（如代谢性疾病）有效治疗的合理药物发现提供了基础。