COUPLED METAL BINDING-PROTEIN FOLDING REACTIONS

大约三分之一的蛋白质与金属离子结合。 在许多生物过程中扮演着至关重要的角色，包括 能量的储存和传递、基因调节和新陈代谢。在……里面 为了让这些蛋白质实现它们的正常功能，它们必须 选择性地结合它们的同源金属离子。这样的识别过程具有 通常被认为是由简单的、热力学的 受控的结合反应。越来越多的证据揭示了 动力学因素的重要性也是如此。金属结合的动力学 交换反应还没有得到广泛的研究。这个 金属结合反应的热力学和动力学似乎是 很大程度上受保留在 去除金属离子后的蛋白质。分子的热力学和动力学 我们将对金属结合、释放和交换反应进行一系列研究 在这些结构特征方面不同的蛋白质。 只有在金属结合条件下才能诱导蛋白质折叠的体系 研究金属结合和蛋白质的独特机会 折叠过程。这类研究将在使用SO- 被称为锌指多肽。蛋白质折叠有待研究的方面 包括不同氨基酸采用的热力学倾向 β-折叠、α-螺旋和其他结构。所获得的数据将是 根据基于结构的热力学分析模型进行解释 已被开发用于研究其他蛋白质折叠和结合 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一个 用于监测锌和其他金属离子的系列多肽传感器 浓度是通过荧光变化实现的。

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.