COUPLED METAL BINDING-PROTEIN FOLDING REACTIONS

偶联金属结合-蛋白质折叠反应

• 批准号: 6107701
• 负责人: Jeremy M. Berg
• 金额: $ 18.31万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-01 至 2000-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6107701
• 关键词: acidity /alkalinity; azurin; calorimetry; chemical binding; chemical kinetics; cobalt; conformation; copper; divalent metal; fluorescent dye /probe; hydropathy; intermolecular interaction; iron; metalloproteins; nickel; nuclear magnetic resonance spectroscopy; peptide chemical synthesis; protein engineering; protein folding; protein structure function; rubredoxins; structural biology; temperature; thermodynamics; zinc

Approximately one third of all protein bind metal ions.Such metalloproteins play crucial roles in a large number of biological processes including the storage and transduction of energy, gene regulation, and metabolism. In order for these proteins to achieve their proper functions, they must selectively bind their cognate metal ions. Such recognition processes have been generally assumed to be governed by simple, thermodynamically controlled, binding reactions. Evidence has been accumulating revealing the importance of kinetic factors as well. The kinetics of metal binding and exchange reactions have not been extensively studied. The thermodynamics and kinetics of metal binding reactions appear to be significantly affected by the degree of structure that remains in the protein upon removal of the metal ion. The thermodynamics and kinetics of metal binding, release, and exchange reactions will be studied for a series of protein that differ in terms of these structural characteristics. Systems for which protein folding is induced only upon metal binding offer unique opportunities for investigating both metal binding and protein folding processes. Such studies will be performed with the use of a so- called zinc finger peptide. Aspects of protein folding to be investigated include the thermodynamic propensities for different amino acids to adopt beta sheet, alpha helix, and other structures. The data obtained will be interpreted in terms of a structure-based thermodynamics analysis model that has been developed for studies of other protein folding and binding reactions. Finally, the ability to modulate metal binding affinity through variations in protein folding energies will be utilized to generate a series of peptide sensors to be used to monitor zinc and other metal ion concentrations via changes in fluorescence.

大约三分之一的蛋白质结合金属离子。 在包括 能量，基因调节和代谢的储存和转导。 在 下令这些蛋白质实现其适当的功能，它们必须 有选择地结合其同源金属离子。 这样的识别过程有 通常假定由简单的热力学控制 受控的结合反应。 证据已经积累了揭示 动力学因素的重要性。 金属结合的动力学 尚未对交换反应进行广泛的研究。 这 金属结合反应的热力学和动力学似乎是 显着受到仍然存在的结构程度的影响 去除金属离子后的蛋白质。 热力学和动力学 将研究一系列的金属结合，释放和交换反应 在这些结构特征方面有所不同的蛋白质。 仅在金属结合报价时诱导蛋白质折叠的系统 研究金属结合和蛋白质的独特机会 折叠过程。 这样的研究将在使用SO-的使用中进行 称为锌指肽。 蛋白质折叠的各个方面要研究 包括用于采用不同氨基酸的热力学倾向 Beta表，Alpha Helix和其他结构。 获得的数据将是 用基于结构的热力学分析模型来解释 已开发用于研究其他蛋白质折叠和结合的研究 反应。 最后，通过调节金属结合亲和力的能力 蛋白质折叠能的变化将用于生成A 一系列用于监测锌和其他金属离子的肽传感器 通过荧光变化的浓度。