MULTINUCLEAR NMR PROBES OF BIOLOGICAL SYSTEMS

生物系统的多核核磁共振探针

• 批准号: 3226148
• 负责人: IAN M ARMITAGE
• 金额: $ 18.38万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-05-01 至 1991-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3226148
• 关键词: Escherichia coli; affinity chromatography; alkaline phosphatase; aminoacid metabolism; aminoacid transport; binding proteins; carbonate dehydratase; carbonic anhydrase inhibitors; catechol methyltransferase; cations; chemical binding; chemical structure function; enzyme structure; enzyme substrate complex; intestines; kidney; laboratory rabbit; laboratory rat; ligands; ligase; lipopolysaccharides; membrane model; membrane permeability; membrane structure; metalloenzyme; metals; methionine; nuclear magnetic resonance spectroscopy; pharmacology

The overall objectives are to develop and apply state-of-the-art NMR techniques to the determination of the solution structural properties of complex biological macromolecules in order to elucidate their mechanism of action. The specific biological systems that will be studied by these methods are metallothioneins and a cyclosporin binding protein, termed cyclophilin. Metallothioneins (MTs): A variety of NMR methods will be used to fully resolve both the structural and metal-ion binding properties of this ubiquitous, metal-ion inducible, low molecular weight, cysteine-rich metal-binding protein isolated from various species in its long evolutionary history (fungus, yeast, invertebrate and mammalian). These very properties assures the ultimate importance of this protein in the cellular regulation of essential (Zn, Cu) and nonessential (Cd, Hg) metal ions. As a result, every effort will be made in these studies to elucidate those aspects of the structure of these different MTs which alter its metal-ion binding properties and to correlate this data with the presumed, but as yet undefined, function of this family of proteins. Cyclophilin is a ubiquitous, low molecular weight (17 Kd) cytosolic protein with high specificity for binding cyclosporin A, a potent immunosuppressant used clinically for the prevention of kidney, liver and heart allograft rejection. The specific objectives will be to use one and two-dimensional 1H NMR methods to elucidate the solution structural properties of cyclophilin, particularly as they relate to the binding site for cyclosporin. Studies such as these are of utmost importance to understanding the role of this protein in the pharmacologic activity of cyclosporin and its active metabolites, and the structure-activity relationships for cyclosporin. The long-term objectives would be to use NMR, in conjunction with other biochemical methods, to characterize the complex of cyclophilin with its intrinsic natural ligand when the latter is isolated. These studies would provide important new insight into the molecular mechanism of action of cyclosporin and the potential physiological significance of cyclophilin in the regulation of cellular metabolism.

总体目标是开发和应用最新的NMR 确定解决方案结构特性的技术 复杂的生物大分子，以阐明其机制 行动。 这些将研究的特定生物系统 方法是金属霉素和环孢蛋白结合蛋白，称为 环磷脂。 金属霉素（MTS）：将使用多种NMR方法来完全 解决此结构和金属离子结合特性 无处不在，金属离子诱导，低分子量，富含半胱氨酸的 从各种物种中分离出的金属结合蛋白 进化史（真菌，酵母，无脊椎动物和哺乳动物）。 这些 非常特性确保该蛋白在 必需（Zn，Cu）和非必需（CD，HG）金属的细胞调节 离子。 结果，将在这些研究中竭尽全力阐明 这些不同MT的结构的那些方面改变了其 金属离子结合特性，并将这些数据与假定的数据相关联 但是尚未定义，这种蛋白质家族的功能。 环肽是一种普遍存在的低分子量（17 kD）胞质蛋白 具有高特异性对于结合环孢菌素A，有效的免疫抑制剂 临床用于预防肾脏，肝脏和心脏同种异体移植 拒绝。 具体目标是使用一个和二维 1H NMR方法阐明了解决方案结构特性 环糖蛋白，特别是当它们与结合位点相关时 环孢菌素。 这样的研究对于 了解该蛋白在药理活性中的作用 环孢菌素及其活性代谢产物及其结构活性 环孢菌素的关系。 长期目标是使用 NMR与其他生化方法结合使用，以表征 当后者为固有的天然配体的复合物时 孤立。 这些研究将为您提供重要的新见解 环孢菌素的作用和电势的分子机制 环粒蛋白在细胞调节中的生理意义 代谢。