Retinoic Acid Receptors: Dynamics, Stability and Folding

视黄酸受体：动力学、稳定性和折叠

• 批准号: 7039105
• 负责人: Michael Schimerlik
• 金额: $ 17.27万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7039105
• 关键词: biochemistry; biological signal transduction; cell differentiation; chemical kinetics; chemical stability; conformation; gene expression; genetic regulation; ligands; liquid chromatography; mass spectrometry; nuclear receptors; protein folding; protein protein interaction; protein purification; protein structure function; retinoate; retinoid binding proteins; thermodynamics

DESCRIPTION (provided by applicant): Retinoid X receptors (RXR) act as homodimers or heterodimers with retinoic acid receptors (RAR) or other nuclear hormone receptors to regulate gene expression. Among the genes regulated by retinoids are those that mediate apoptosis, cholesterol absorption and efflux, expression of xenobiotic metabolizing enzymes and the oncogene v-Erb A. Because of the important roles that RXR and RAR play in growth, development and cellular differentiation retinoids are of interest as chemopreventative and chemotherapeutic agents. They are also used in the treatment of acne, psoriasis and precancerous lesions. Although X-ray structures of several receptors for retinoids have been determined, information concerning the dynamics and stability of these proteins and the effects of ligands on these properties is lacking. This proposal will use mass spectrometry in conjunction with hydrogen/deuterium exchange to (1) examine the dynamics of the RARgamma and RXRalpha ligand binding domains and the effects of ligands and protein quaternary structure on solvent accessibility of structural elements, (2) probe the kinetic mechanism by which these proteins fold and (3) analyze conformational changes by differential protease sensitivity. Thermodynamic and kinetic studies are also proposed to examine the energetics and mechanisms of folding. These studies give information that complements the static, structural data, forms the foundation for future work to characterize the interactions of these proteins with other components of their signal transduction pathways and provides new data that may aid in the design of more effective drugs that act on receptors for retinoids.

描述（由申请人提供）：类维生素A X受体（RXR）与视黄酸受体（RAR）或其他核激素受体作为同源二聚体或异源二聚体发挥作用，以调节基因表达。类维生素A调节的基因包括介导细胞凋亡、胆固醇吸收和流出、异生物质代谢酶的表达和致癌基因v-Erb A。由于RXR和RAR在生长、发育和细胞分化中起重要作用，类维生素A作为化学预防剂和化学治疗剂受到关注。 它们也用于治疗痤疮、牛皮癣和癌前病变。 虽然已经确定了几种类维生素A受体的X射线结构，但缺乏有关这些蛋白质的动力学和稳定性以及配体对这些性质的影响的信息。该提案将使用质谱法结合氢/氘交换来（1）检查RAR γ和RXR α配体结合结构域的动力学以及配体和蛋白质四级结构对结构元件的溶剂可及性的影响，（2）探测这些蛋白质折叠的动力学机制，以及（3）通过差异蛋白酶敏感性分析构象变化。热力学和动力学的研究也提出了审查折叠的能量和机制。这些研究提供的信息补充了静态的结构数据，为未来的工作奠定了基础，以表征这些蛋白质与其信号转导途径的其他组分的相互作用，并提供了新的数据，可能有助于设计更有效的药物作用于类维生素A受体。