PROTEIN-PROSTHETIC GROUP STABILITY, DYNAMICS & FUNCTION

蛋白质修复基团的稳定性、动力学

• 批准号: 6180382
• 负责人: A. JOSHUA WAND
• 金额: $ 38.58万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-01-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6180382
• 关键词: binding proteins; calorimetry; chemical stability; computer simulation; cytochrome b; cytochrome c; electron transport; enzyme complex; flavins; flavodoxin; heme; hemoprotein structure; intermolecular interaction; molecular dynamics; nuclear magnetic resonance spectroscopy; oxidation reduction reaction; protein engineering; protein folding; protein structure function; thermodynamics

This proposal focuses on several important issues underlying the folding, stability, dynamics and functional state dependence of prosthetic group binding proteins. Our goal is to contribute to the understanding of the contribution of structure and dynamics to the functional competence of flavin and heme proteins, such as those involved in electron transfer. Electron transfer reactions between proteins control the generation, flow and use of energy in biological systems and one third of known enzymes are involved in oxidation and reduction. Accordingly, redox dysfunctional is directly involved with a variety of fetal or debilitating syndromes. We are most interested in exploring why many b-heme and flavin binding proteins display considerate structure and stability in their corresponding apostates and how binding modifies both the prosthetic group and the protein. Specifically, we propose to characterize the structural cooperativity of three apoproteins using hydrogen exchange, NMR relaxation, and a variety of structural NMR methods; we propose to directly quantify the redox- state dependence of dynamics of several holoproteins in an effort to understand the nature of the prosthetic group-protein interface; we propose to use our accumulated knowledge to continue the design and engineering of a series of minimalist proteins, "maquettes", with the ultimate aim of constructing functionally useful and catalytically active mini-proteins which might serve as advanced pharmaceutics and (bio)chemical reagents; and we propose to characterize the complex between cytochrome c and its binding domain in Apaf-1, an interaction that is a key event in apoptosis, programmed cell death. Each of these specific aims will require the extensive use of multi-nuclear and multi- dimensional NMR spectroscopy and will take advantage o f recent methodological developments and novel capabilities, particularly NMR relaxation methods and a high pressure NMR capability.

该提案重点介绍了有关的几个重要问题 折叠，稳定性，动力学和功能状态依赖性 假体组结合蛋白。我们的目标是为 理解结构和动态对 黄素和血红素蛋白的功能能力，例如 参与电子传输。电子转移反应 蛋白质控制生物学中能量的产生，流动和使用 系统和三分之一的已知酶参与氧化和 减少。因此，氧化还原功能失调直接与 多种胎儿或衰弱的综合征。我们最感兴趣 探索为什么许多B-血红素和黄素结合蛋白显示 其相应叛教者的体贴结构和稳定性 结合如何修饰假体组和蛋白质。 具体而言，我们建议表征 三种使用氢交换，NMR松弛和多种多样的载脂蛋白 结构NMR方法；我们建议直接量化氧化还原 几种多种蛋白质的动态依赖性 了解假肢组蛋白接口的性质；我们 建议利用我们积累的知识继续设计和 一系列极简蛋白的工程，“ maquettes”，带有 在功能上有用和催化的最终目标 活跃的迷你蛋白可能是高级药物和 （生物）化学试剂；我们建议表征复合体 在APAF-1中的细胞色素C及其结合域之间 这是细胞凋亡，程序性细胞死亡的关键事件。每个 具体目标将需要广泛使用多核和多核 尺寸NMR光谱法，将利用最近的优势 方法论发展和新功能，尤其是NMR 松弛方法和高压NMR能力。