T4 LYSOZYME AS A MODEL FOR PROTEIN STABILITY

T4 溶菌酶作为蛋白质稳定性的模型

• 批准号: 6180747
• 负责人: Frederick W. Dahlquist
• 金额: $ 20.64万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6180747
• 关键词: atomic force microscopy; bacteriophage T4; conformation; crosslink; hydropathy; lysozyme; mass spectrometry; mathematical model; nuclear magnetic resonance spectroscopy; physical model; protein folding; protein structure function; recombinant proteins; stop flow technique

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.

描述：本提案旨在获得更好的 了解蛋白质结构、稳定性和蛋白质之间的关系 使用 T4 溶菌酶作为模型系统进行动力学研究。 该提案提出了四点 具体目标：1）连接折叠和展开的折叠路径 各国将受到调查，特别强调结构和 出现的早期动力学中间态和平衡态的动力学 与之相关。 描述了监测相对比率的方法 使用准分子探针遇到多肽链的各个部分。 2）序列不同部分之间的耦合机制，即 在野生型蛋白质中作为一个明显的单位折叠，但在 某些突变体。 将采用脉冲氢交换方法 定义蛋白质不同部分达到天然状态的速率 结构。; 3) 水和小疏水配体的可及性 将检查 T4 溶菌酶核心的空腔。 4）重大努力 将着手开发观察和解释所需的方法 应用机械力的效果旨在展开小 附着在原子力显微镜尖端的蛋白质组装体。 这些类型的测量将为能源提供新的视角 蛋白质景观。