PROTEIN-PROSTHETIC GROUP STABILITY, DYNAMICS & FUNCTION

蛋白质修复基团的稳定性、动力学

• 批准号: 6783928
• 负责人: A. JOSHUA WAND
• 金额: $ 13.21万
• 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 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6783928
• 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.

这项提案侧重于以下几个重要问题 折叠、稳定性、动力学和功能状态依赖性 假体基团结合蛋白。我们的目标是为 理解结构和动力学对经济增长的贡献 黄素和血红素蛋白的功能活性，如 参与电子转移的。分子间的电子转移反应 蛋白质控制着生物中能量的产生、流动和使用 系统和已知的三分之一的酶参与氧化和 还原。因此，氧化还原功能障碍直接与 各种胎儿综合症或衰弱综合症。我们最感兴趣的是 探索为什么许多b-血红素和黄素结合蛋白显示 在它们相应的变节和变节中体贴的结构和稳定性 结合如何改变假体基团和蛋白质。 具体地说，我们建议刻画结构协作性 三种脱辅基蛋白采用氢交换、核磁共振弛豫等多种方法 结构核磁共振方法；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关系 了解假体基团-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和 设计了一系列极简主义蛋白质，“maquettes”，与 最终目的是构建功能上有用的催化结构 活性微蛋白可能作为先进的药剂学和 (生物)化学试剂；我们建议对该络合物进行表征 APAF-1中细胞色素c与其结合域之间的相互作用 这是细胞凋亡中的一个关键事件，即程序性细胞死亡。这其中的每一个 具体目标将要求广泛使用多核和多核技术。 三维核磁共振波谱，并将利用最近的 方法的发展和新的能力，特别是核磁共振 松弛方法和高压核磁共振能力。