COUPLED METAL BINDING-PROTEIN FOLDING REACTIONS

偶联金属结合-蛋白质折叠反应

• 批准号: 6316667
• 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: 2000
• 资助国家: 美国
• 起止时间: 2000-06-01 至 2002-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6316667
• 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。此类金属蛋白质 在许多生物过程中发挥着至关重要的作用，包括 能量的储存和传导、基因调控和新陈代谢。 在 为了使这些蛋白质实现其适当的功能，它们必须 选择性地结合它们的同源金属离子。 这样的识别过程有 通常被认为是由简单的热力学控制 受控的结合反应。 证据不断积累 动力学因素的重要性也是如此。 金属结合动力学 和交换反应尚未得到广泛研究。 这 金属结合反应的热力学和动力学似乎是 受残留结构程度的显着影响 去除金属离子后的蛋白质。 热力学和动力学 将研究一系列金属结合、释放和交换反应 这些结构特征不同的蛋白质。 仅在金属结合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。 此类研究将使用以下方法进行： 称为锌指肽。 待研究的蛋白质折叠方面 包括不同氨基酸采用的热力学倾向 β片层、α螺旋和其他结构。 获得的数据将是 根据基于结构的热力学分析模型进行解释 是为研究其他蛋白质折叠和结合而开发的 反应。 最后，通过调节金属结合亲和力的能力 蛋白质折叠能量的变化将被用来产生 用于监测锌和其他金属离子的系列肽传感器 通过荧光的变化来确定浓度。