Structure and Functions of Steroid Hormone Receptors

类固醇激素受体的结构和功能

• 批准号: 7414039
• 负责人: H Eric XU
• 金额: $ 40.34万
• 依托单位: VAN ANDEL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7414039
• 关键词: Adverse effects; Affinity; Agonist; Amino Acid Receptors; Amino Acid Sequence; Androgen Receptor; Anti-Inflammatory Agents; Anti-inflammatory; Asthma; Autoimmune Diseases; Binding; Biological Assay; Body Fluids; C-terminal; Cells; Childhood Acute Lymphocytic Leukemia; Clinical; Collaborations; Complement; Complex; Condition; Consensus; Corticosterone; Crystallization; DNA; DNA Binding Domain; DNA-Protein Interaction; Data; Dexamethasone; Disease; Drug Delivery Systems; Elements; Estrogen Receptors; Family; Figs - dietary; Fluticasone propionate; Gene Targeting; Genetic Transcription; Glucocorticoid Receptor; Glucocorticoids; Goals; Heart failure; Homeostasis; Hormones; Human; Hydrocortisone; Hypertension; Immune; Inflammation; Inflammatory Response; Ketones; Knowledge; Ligand Binding; Ligand Binding Domain; Ligands; Mediating; Medical; Metabolic; Metabolism; Mineralocorticoid Receptor; Molecular; Mutation; N-terminal; National Institute of Diabetes and Digestive and Kidney Diseases; Numbers; Pharmacologic Substance; Physiological; Physiology; Positioning Attribute; Progesterone Receptors; Proteins; RU-486; Range; Receptor Activation; Regulation; Reporting; Reproduction; Research; Research Personnel; Response Elements; Role; Sexual Development; Signal Transduction; Site; Site-Directed Mutagenesis; Sodium; Solutions; Spironolactone; Steroid Receptors; Structure; Structure-Activity Relationship; TATA-Box Binding Protein; Testing; Transactivation; United States National Institutes of Health; Work; base; blood pressure regulation; cofactor; cortivazol; cross reactivity; deacylcortivazol; design; drug discovery; eplerenone; hypertensive heart disease; insight; intermolecular interaction; leukemia; member; next generation; promoter; receptor; steroid hormone receptor

DESCRIPTION (provided by applicant): The long-term objective of this study is to reveal the structural mechanisms of hormone actions mediated by the glucocorticoid receptor (GR) and the mineralococorticoid receptor (MR). GR and MR are members of the steroid hormone receptor family that regulate broad spectrum of human physiology ranging from immune/inflammatory responses to metabolic homeostasis and control of blood pressure. Importantly, both GR and MR are well-established drug targets, with a number of their ligands currently being used to treat diverse diseases including inflammation and hypertension. GR and MR mediate their actions through hormone binding to their C-terminal ligand-binding domain (LBD) and promoter binding of target genes to their middle DMA-binding domain (DBD). Both receptors also contain a potent N-terminal activation domain (AF-1). Until recently, only structures of the isolated DBD or the isolated GR LBD with a single agonist (dexamethasone) had been solved. Clearly, this limited amount of structural information is a serious deficiency considering the importance of GR and MR in normal physiology and in disease. In this study, we propose to fill in that knowledge gap by solving the crystal structures of various GR and MR complexes. Our specific aims are focused on crystallization and structural determination of 1) the MR LBD, 2) the GR LBD in complex with cortivazol, a potent anti-inflammatory ligand that binds to GR with 40-fold higher affinity than dexamethasone, 3) a MR DBD-LBD fragment bound to the MR consensus DNA site, and 4) the GR AF-1-DBD/DNA complex that contains the GR AF-1 domain and its DBD. The hypothesis for our specific aims is that the molecular interactions (protein-protein, protein-DNA, and protein-hormone) observed in these crystal structures will be crucial for mechanistic understanding of the hormone actions of GR and MR. After structural determination, we will identify key structural elements by scrutinizing and analyzing the structures, and we will collaborate closely with Stoney Simons (NIDDK), Brad Thompson, and Raj Kumar (UTMB, Galveston), in site-directed mutagenesis and cell-based transcriptional assays to assess the functional significance of the key features identified. Significance: The structural information generated in this application will significantly enhance our understanding of the molecular mechanisms of hormone actions by GR and MR, and should serve as rational templates for drug discovery that targets these 2 receptors for the treatment of asthma, hypertension, and heart disease.

描述（由申请人提供）：本研究的长期目标是揭示糖皮质激素受体（GR）和盐皮质激素受体（MR）介导的激素作用的结构机制。GR和MR是类固醇激素受体家族的成员，其调节从免疫/炎症反应到代谢稳态和血压控制的广谱人类生理学。重要的是，GR和MR都是公认的药物靶点，它们的许多配体目前被用于治疗包括炎症和高血压在内的多种疾病。GR和MR通过激素与其C-末端配体结合结构域（LBD）的结合以及靶基因与其中间DMA-结合结构域（DBD）的启动子结合来介导其作用。这两种受体还含有一个有效的N末端激活结构域（AF-1）。直到最近，只有分离的DBD或分离的GR LBD与单一激动剂（地塞米松）的结构已经解决。显然，考虑到GR和MR在正常生理和疾病中的重要性，这种有限的结构信息是一个严重的缺陷。在这项研究中，我们建议通过解决各种GR和MR复合物的晶体结构来填补这一知识空白。我们的具体目标集中在以下各项的结晶和结构测定：1）MR LBD，2）GR LBD与可替唑（一种与GR结合的亲和力比地塞米松高40倍的强效抗炎配体）复合，3）与MR共有DNA位点结合的MR DBD-LBD片段，以及4）含有GR AF-1结构域及其DBD的GR AF-1-DBD/DNA复合物。我们的特定目标的假设是，在这些晶体结构中观察到的蛋白质-蛋白质、蛋白质-DNA和蛋白质-激素）对于GR和MR的激素作用的机理理解将是至关重要的。在结构确定之后，我们将通过仔细检查和分析结构来识别关键结构元件，并且我们将与Stoney Simons（NIDDK）、布拉德汤普森、和Raj Kumar（UTMB，加尔维斯顿），在定点诱变和基于细胞的转录测定中评估所鉴定的关键特征的功能意义。重要性：本申请中产生的结构信息将显著增强我们对GR和MR激素作用的分子机制的理解，并应作为药物发现的合理模板，靶向这2种受体用于治疗哮喘，高血压和心脏病。