Hydrogen exchange, proteins, folding, and misfolding

氢交换、蛋白质、折叠和错误折叠

• 批准号: 7033307
• 负责人: S. Walter ENGLANDER
• 金额: $ 38.05万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-12-01 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7033307
• 关键词: Staphylococcus; alpha synuclein; amyloid proteins; bacterial proteins; biophysics; chemical kinetics; chemical models; cytochrome c; high throughput technology; hydrogen; mass spectrometry; nuclear magnetic resonance spectroscopy; nuclease; protein folding; protein structure function; thermodynamics

DESCRIPTION (provided by applicant): Work supported by this grant over the years helped to develop the hydrogen exchange knowledge and methods that are now widely exploited in studies of protein structure and function. Progress during this grant period has uncovered a fundamental but previously unsuspected protein property. It appears that proteins are composed of a small number of cooperative unfolding/refolding units, called foldons, and that these units form and assemble in a pathway sequence through distinct intermediate forms to progressively construct the native protein. The present application proposes to further test this hypothesis in cytochrome c. Also, the generality of this behavior will be tested with other major protein models, using staphylococcal nuclease initially and then other model proteins to be selected according to specified criteria. In other work to continue the development of hydrogen exchange knowledge and its use for understanding protein dynamics and signal transfer, designed mutational perturbations will be used to study the local fluctuation mechanisms of proteins and the internal protein pathways that mediate more distant changes. All of this work will use special hydrogen exchange methods that we have developed and will depend on NMR analysis. Related hydrogen exchange experiments that depend on mass spectroscopic analysis, together with other methods, will be used to study amyloid structures and their precursor forms. The focus will be on alpha-synuclein initially (Parkinson's Disease) and then Alpha/Beta (Alzheimer's Disease) and will use high throughput methods among others, looking toward drug discovery efforts. The research described here aims to understand the functioning of the protein machinery of biology, both in health and in disease. This work has uncovered and is continuing to study a previously unsuspected property of protein molecules, their cooperative foldon substructure, which appears to explain how proteins fold to their active form and how they perform some other functions. The knowledge gained and the special methods developed are also being used to study the protein misfolding mechanisms that lead to devastating human and animal neurodegenerative pathologies such as Parkinson's and Alzheimer's disease and bovine spongiform encephalopathy (mad cow disease).

描述（由申请人提供）：多年来这项资助支持的工作有助于发展氢交换知识和方法，这些知识和方法现在广泛用于蛋白质结构和功能的研究。在此资助期间取得的进展揭示了一种基本但之前未被怀疑的蛋白质特性。蛋白质似乎是由少量协作的展开/重折叠单元（称为折叠单元）组成，并且这些单元通过不同的中间形式以路径序列形成和组装，以逐步构建天然蛋白质。本申请提出在细胞色素c中进一步检验该假设。此外，将使用其他主要蛋白质模型来测试这种行为的普遍性，首先使用葡萄球菌核酸酶，然后根据指定标准选择其他模型蛋白质。在继续发展氢交换知识及其用于理解蛋白质动力学和信号传递的其他工作中，设计的突变扰动将用于研究蛋白质的局部波动机制和介导更远距离变化的内部蛋白质途径。所有这些工作都将使用我们开发的特殊氢交换方法，并依赖于核磁共振分析。依赖于质谱分析的相关氢交换实验以及其他方法将用于研究淀粉样蛋白结构及其前体形式。重点将首先集中在 α-突触核蛋白（帕金森病），然后是 Alpha/Beta（阿尔茨海默病），并将使用高通量方法等，着眼于药物发现工作。这里描述的研究旨在了解生物学蛋白质机制在健康和疾病中的功能。这项工作已经发现并正在继续研究蛋白质分子的一个以前未曾怀疑过的特性，即它们的协同折叠子结构，这似乎解释了蛋白质如何折叠成其活性形式以及它们如何执行一些其他功能。所获得的知识和开发的特殊方法也被用于研究蛋白质错误折叠机制，这些机制会导致毁灭性的人类和动物神经退行性疾病，例如帕金森病和阿尔茨海默病以及牛海绵状脑病（疯牛病）。