Retinoic Acid Receptors: Dynamics, Stability and Folding

视黄酸受体：动力学、稳定性和折叠

• 批准号: 6617171
• 负责人: Michael Schimerlik
• 金额: $ 17.69万
• 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/6617171
• 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(申请人提供)：维甲酸X受体(RXR)与维甲酸受体(RAR)或其他核激素受体一起作为同源二聚体或异源二聚体来调节基因表达。在维甲酸调控的基因中，包括介导细胞凋亡、胆固醇吸收和排出的基因、异源代谢酶的表达和癌基因v-erb A。由于RXR和RAR在生长、发育和细胞分化中发挥重要作用，维甲酸作为化学预防和化疗药物引起了人们的兴趣。它们还用于治疗痤疮、牛皮癣和癌前病变。虽然已经确定了几种维甲酸受体的X射线结构，但关于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还很少。这一建议将结合氢/氢交换使用质谱学来(1)检测RARGamma和RXRpha配体结合域的动力学以及配体和蛋白质四级结构对结构元素溶剂可及性的影响，(2)探索这些蛋白质折叠的动力学机制，(3)通过差异蛋白酶的敏感性来分析构象变化。还建议进行热力学和动力学研究，以考察折叠的能量学和机理。这些研究提供了补充静态结构数据的信息，为未来研究这些蛋白质与其信号转导途径的其他成分的相互作用奠定了基础，并提供了新的数据，可能有助于设计更有效的作用于维甲酸受体的药物。