T4 LYSOZYME AS A MODEL FOR PROTEIN STABILITY

描述：本建议旨在获得更好的 了解蛋白质结构、稳定性和 动力学使用T4溶菌酶作为模型系统。 该提案提出了四个 具体目标：1）连接折叠和展开的折叠路径 国家将进行调查，特别强调结构和 早期动力学中间体和平衡态的动力学， 描述了监测其相对速率的方法， 用受激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， 在野生型蛋白中折叠为表观单位，但在 某些变种人 将采用脉冲氢交换方法， 定义了蛋白质不同部分获得类似天然蛋白质的速率， 结构。3)水和小的疏水配体的可及性 将检查T4溶菌酶核心中的空腔。4)一项重大努力 将着手开发必要的方法来观察和解释 设计用于展开小的机械力的应用的效果 附着在原子力显微镜尖端的蛋白质组合体。 这些类型的测量将提供一个新的视角的能源 蛋白质景观

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.