New assay method for pinpointing structural features in amyloid oligomer formation

精确定位淀粉样蛋白寡聚体形成结构特征的新测定方法

• 批准号: 10214240
• 负责人: CORIE Y RALSTON
• 金额: $ 55.87万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-15 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10214240
• 关键词: Affect; Alzheimer' s Disease; Alzheimer' s disease therapeutic; Amino Acid Sequence; Amyloid; Amyloid Fibrils; Amyloid Proteins; Amyloid beta-Protein; Binding; Biological Assay; Biomedical Research; Charge; Clinical Trials; Complex; Data; Development; Disease; Disease Progression; Failure; Future; Goals; Grant; Hour; Hydroxyl Radical; Impaired cognition; Investigation; Lead; Mass Spectrum Analysis; Methodology; Methods; Molecular; Molecular Conformation; Mutation; Noise; Pathogenesis; Pathway interactions; Peptides; Process; Protein Conformation; Protein Dynamics; Proteins; Public Health; Research Personnel; Roentgen Rays; Role; Sampling; Signal Transduction; Solvents; Structure; System; Techniques; Testing; Therapeutic; Time; Toxic effect; Variant; Work; aggregation pathway; amyloid peptide; amyloid precursor protein processing; base; brain tissue; design; drug discovery; familial Alzheimer disease; instrument; ion mobility; millisecond; misfolded protein; neurotoxicity; new technology; novel therapeutics; peptide P3; protein aminoacid sequence; stoichiometry; therapeutic target; tool

Summary The goal of this project is to provide a significant new technology for biomedical research, particularly for researchers studying proteins associated with the progression of Alzheimer's Disease (AD). The majority of efforts to develop therapeutics for AD, most of which have targeted Aβ fibrils, have failed. A growing body of evidence now suggests that oligomeric forms of Aβ, which are intermediates that form during the aggregation process, may be the culprits in disease progression. However, in order to design therapeutics targeting toxic oligomeric species or other altered protein forms, we first require structural information on those species. Here, we propose a new structural method to characterize oligomeric intermediates in aggregation-prone systems, and to test the new methodology on specific amyloid peptides implicated in AD. This work will develop a new structural assay which synergistically combines local and global structural mass spectrometry-based methods. The hydroxyl radical solvent accessibility method reveals local residue-specific structural changes that occur as a function of conformational changes or complex formation, while the native mass spectrometry method reveals global structural information on oligomer size and stoichiometry. These methods will be applied to the study of the amyloid peptides in the familial forms of AD to determine how the oligomeric states of these peptides differ from the wildtype. The new assay will also be used to characterize the interaction between amyloid peptides and the p3 peptide, which is twice as abundant as the more well-studied Aβ, fibrilzes faster than Ab, but can mitigate Ab toxicity. Finally, we will relate these intermediate structural forms to toxicity. These results will be of immense therapeutic value for AD, and will lay the groundwork for future structural investigations of difficult-to-study intermediate forms in a wide range of diseases caused by the aggregation or misfolding of proteins.

概括 该项目的目的是为生物医学研究提供重要的新技术， 特别是对于研究与阿尔茨海默氏症进展相关的蛋白质的研究人员 疾病（AD）。开发广告疗法的大多数努力，其中大多数 靶向的Aβ原纤维失败了。现在越来越多的证据表明低聚形式 在聚集过程中形成的中间体的Aβ可能是罪魁祸首 疾病进展。但是，为了设计靶向有毒寡聚物种的理论 或其他改变的蛋白质形式，我们首先需要有关这些物种的结构信息。在这里，我们 提出一种新的结构方法，以表征易聚合中的低聚中间体 系统，并测试有关AD中实施的特定淀粉样蛋白宠物的新方法。这 工作将开发出一种新的结构测定 基于结构质谱的方法。羟基自由基溶剂可及性法 揭示了当地居住的特定结构变化，这些变化是构象的函数 变化或复杂的形成，而天然质谱法则揭示了全局 关于低聚物大小和化学计量的结构信息。这些方法将应用于 研究淀粉样蛋白在AD的家族形式中的研究，以确定寡聚状态如何 这些宠物与野生型不同。新测定也将用于表征 淀粉样蛋白的淀粉样蛋白与P3 Pepperide之间的相互作用，其丰富的两倍 研究的Aβ比AB更快，纤维纤维更快，但可以减轻AB的毒性。最后，我们将联系 这些中间结构形式对毒性。这些结果将是巨大的治疗 对AD的价值，并将为未来难以研究的结构调查奠定基础 由聚集或错误折叠引起的多种疾病中的中间形式 蛋白质。