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.

该提案侧重于几个重要问题， 折叠，稳定性，动力学和功能状态依赖性 辅基结合蛋白。我们的目标是为 理解结构和动力学对 黄素和血红素蛋白质的功能能力，如那些 参与电子转移。电子转移反应 蛋白质控制着生物体内能量的产生、流动和使用。 系统和三分之一的已知酶参与氧化， 还原因此，氧化还原功能障碍直接涉及细胞的生长。 各种胎儿或衰弱综合征。我们最感兴趣 探索为什么许多血红素和黄素结合蛋白 在他们相应的叛教者中， 结合如何改变辅基和蛋白质。 具体地说，我们建议表征结构的协同性， 三个载脂蛋白使用氢交换，核磁共振弛豫，和各种 结构NMR方法;我们建议直接量化氧化还原- 几种全蛋白动力学的状态依赖性， 了解辅基-蛋白质界面的性质;我们 建议利用我们积累的知识继续设计， 一系列极简蛋白质的工程，“maquettes”， 最终目的是建设功能有用和催化 活性微蛋白可能作为高级药物， （生物）化学试剂;我们建议表征复合物 Apaf-1中细胞色素c与其结合结构域之间的相互作用 这是细胞凋亡中的一个关键事件，程序性细胞死亡。这一切成功都 具体目标将需要广泛使用多核武器和多 三维NMR光谱，并将利用最近的 方法的发展和新的能力，特别是NMR 弛豫方法和高压NMR能力。