T4 LYSOZYME AS A MODEL FOR PROTEIN STABILITY

描述：这项建议旨在获得更好的 理解蛋白质结构、稳定性和蛋白质之间的关系 以T4溶菌酶为模型系统进行动力学研究。该提案提出了四个方面 具体目标：1)连接折叠和展开的折叠通道 将对各州进行调查，特别强调结构和 早期动力学中间体的动力学和出现的平衡态 与之相关的。描述了监控相对增长率的方法 用准分子探针使多肽链的不同部分相遇。 2)序列不同部分之间的耦合机制，即 在野生型蛋白中折叠为一个明显的单位，但在 某些变种人。将采用脉冲氢交换方法来 定义蛋白质的不同部分达到天然状态的速率 结构；3)水和小分子疏水配体的可及性 将检查T4溶菌酶核心中的空洞。4)主要工作 将致力于发展观察和解释所需的方法 机械力的应用效果设计成展开小 附着在原子力显微镜顶端的蛋白质集合。 这些类型的测量将提供对能量的新视角 蛋白质的景观。

DESCRIPTION: This proposal is directed towards obtaining a better understanding of the relationships between protein structure, stability and dynamics using T4 lysozyme as a model system. The proposal presents four specific aims: 1) The folding pathway connecting the folded and unfolded states will be investigated, with specific emphasis on the structure and dynamics of an early kinetic intermediate and equilibrium states that appear related to it. Approaches are described to monitor the relative rates of encounter of various parts of the polypeptide chain using an excimer probe.; 2) The mechanism of coupling between different parts of the sequence, that fold as an apparent unit in the wild-type protein but become uncoupled in certain mutants. Pulsed hydrogen exchange methods will be employed to define the rates at which different parts of the protein attain native-like structure.; 3) The accessibility of water and small hydrophobic ligands to cavities in the core of T4 lysozyme will be examined.; 4) A major effort will be undertaken to develop the methods necessary to observe and interpret the effects of the application of mechanical forces designed to unfold small assemblies of proteins attached to the tip of an atomic force microscope. These types of measurements will provide a new perspective of the energy landscape of proteins.