Protein structure and function by hydrogen exchange mass spectrometry analysis

氢交换质谱分析蛋白质结构和功能

• 批准号: 8755781
• 负责人: S. Walter ENGLANDER
• 金额: $ 32.26万
• 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 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8755781
• 关键词: Aging-Related Process; Alleles; Alzheimer' s Disease; Amides; Amino Acids; Amyloid; Antibodies; Apolipoprotein A-I; Apolipoproteins; Area; Autoantibodies; Autoimmune Diseases; Autoimmunity; Behavior; Binding; Binding Sites; Biological Process; Biology; Cardiovascular Diseases; Cells; Complex; Development; Disease; Epitopes; Functional disorder; Funding; Gerda brand of difluprednate; Grant; High Density Lipoproteins; Hydrogen; Kinetics; Knowledge; Label; Laboratories; Learning; Lipid Binding; Lipids; Mass Spectrum Analysis; Measurement; Measures; Membrane; Metalloproteases; Methodology; Methods; Motion; Nucleotides; Process; Proline; Property; Protein Isoforms; Proteins; Resolution; Risk; Structural Protein; Structure; System; Technology; Therapeutic; Therapeutic antibodies; Thermodynamics; Thrombotic Thrombocytopenic Purpura; United States National Institutes of Health; Work; amyloid formation; apolipoprotein E-3; apolipoprotein E-4; biophysical properties; genetic risk factor; lipid metabolism; member; protein aggregation; protein complex; protein function; protein protein interaction; protein structure function; public health relevance; research study

DESCRIPTION (provided by applicant): The major challenge for the protein chemist is to explain how proteins implement their limited repertoire of structural and biophysical properties to produce their myriad functions. Unlike any other laboratory method, hydrogen exchange (HX) behavior encodes detailed quantitative information, at amino acid resolution, on the biophysical factors that produce protein function -- structure, structure change, interactions, dynamics, and energetics. Previous versions of this NIH grant have been instrumental in developing the HX field by using a variety of NMR methods, but routine NMR analysis is limited to small highly soluble proteins in substantial quantity and high concentration. A developing hydrogen exchange - mass spectrometry technology (HX MS) promises to extend this proven capability to the larger and more complex protein systems that make biology work while requiring only picomoles of protein at sub-micromolar concentrations. This application proposes to perfect the HX MS technology and apply it in studies of important protein systems. They are: 1) apolipoproteins E3 and E4, when lipid- free and lipid-bound, important for cardiovascular and Alzheimer's diseases; 2) the definition of autoimmune antibody epitopes in acquired TTP disease, relevant to antibody therapeutics and to protein interactions more generally; 3) the Hsp104 protein disaggregase, relevant to the control of protein aggregation and amyloid processing. The full development of HX MS and its demonstration with difficult but centrally important protein systems will provide a uniquely powerful methodology that is widely applicable to the study of protein structure - function problems.

描述（由申请人提供）：蛋白质化学家的主要挑战是解释蛋白质如何实施其结构和生物物理特性的有限曲目 产生他们无数的功能。与任何其他实验室方法不同，氢交换（HX）行为在氨基酸分辨率上编码了产生蛋白质功能的生物物理因子 - 结构，结构变化，相互作用，动力学和能量学。该NIH赠款的先前版本通过使用多种NMR方法对开发HX领域的作用有助于，但常规的NMR分析仅限于大量和高浓度的小型高度溶性蛋白。开发的氢交换 - 质谱技术（HX MS）有望将这种可靠的能力扩展到更大，更复杂的蛋白质系统，这些系统使生物学起作用，同时仅需要蛋白质蛋白质蛋白在亚微摩尔浓度下。该应用建议完善HX MS技术，并将其应用于重要的蛋白质系统的研究。它们是：1）载脂蛋白E3和E4，当无脂质和脂质结合时，对心血管和阿尔茨海默氏病很重要； 2）在获得的TTP疾病中的自身免疫性抗体表位的定义与抗体疗法有关，并且更普遍地与蛋白质相互作用有关； 3）HSP104蛋白分解酶，与控制蛋白质聚集和淀粉样蛋白加工有关。 HX MS的全面发展及其具有困难但中心重要的蛋白质系统的演示将提供一种独特的强大方法，该方法广泛适用于蛋白质结构 - 功能问题的研究。