Copper-Induced Amyloidosis Studied by Mass Spectrometry

通过质谱研究铜诱导的淀粉样变性

基本信息

批准号：
9382123
负责人：
RICHARD W VACHET
金额：
$ 30.97万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-07-01 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9382123
关键词：
Allosteric Site Alzheimer&apos s Disease Amino Acids Amyloid Amyloid Fibrils Amyloid Proteins Amyloidosis Binding Binding Sites Complement Copper Data Deuterium Dialysis procedure Disease Gases Grant Human Hydrogen Individual Isomerism Label Mass Spectrum Analysis Measurement Methods Modeling Molecular Nature Non-Insulin-Dependent Diabetes Mellitus Outcomes Research Parkinson Disease Pathogenicity Pathway interactions Phase Play Process Proteins Reaction Reagent Reporting Research Role Side Site Structural Protein System Techniques Testing Therapeutic Variant Vertebral column Work amyloid fibril formation amyloid formation base cis trans isomerization conformer experimental study fascinate human disease improved inhibitor/antagonist insight ion mobility monomer novel novel strategies protein structure small molecule synergism therapeutic development tool

项目摘要

Project Summary Several human proteins are known to form amyloid fibrils, and these fibrils are associated with several devastating diseases, including Alzheimer’s, Parkinson’s, type II diabetes, and dialysis-related amyloidosis (DRA). One of these proteins, human β-2- microglobulin (β2m), can form amyloid fibrils in the presence of Cu(II), leading to DRA. While many general aspects of amyloid formation are understood, molecular-level information about the early stages of amyloid forming reactions is only beginning to be revealed. This information, though, is critical for the rational development of therapeutics against amyloid diseases. Previous work from our group has developed and applied new covalent labeling (CL) techniques together with mass spectrometry (MS) to structurally characterize the pre-amyloid oligomers of β2m. The insight gained from these new tools has helped identify small molecules that can inhibit β2m amyloid formation. Understanding the molecular basis of these inhibitors is important for improving them, and we will advance new MS-based approaches to gain this understanding. MS is emerging as a powerful tool for studying the structures of proteins and protein oligomers, but most often individual MS-based techniques are used to gather the desired information. We propose the combination of MS-based structural tools that together will provide synergistic information to characterize the molecular basis of β2m amyloid inhibitors. Specifically, we will explore the combination of CL/MS and hydrogen/deuterium exchange (HDX)/MS to give a more detailed picture of the structural changes undergone by β2m and its oligomers. We will also establish an HDX-enhanced ion mobility method to characterize isomeric oligomers that are populated in the presence of certain inhibitors. Our research on β2m amyloid formation so far has revealed how Cu(II) partially unfolds β2m, allowing it to form distinct oligomers before mature fibrils are produced. We have also begun to study inhibitors of β2m amyloid formation. The combination of MS-based methods proposed here seek to reveal the molecular basis of inhibitors of β2m amyloid formation, including characterization of isomeric oligomers. A specific outcome of this research will be a better molecular-level understanding of how to inhibit β2m amyloid formation, which will lead to therapeutics against DRA. The techniques developed in this work will be generally applicable to other amyloid systems.

项目概要已知几种人类蛋白质可形成淀粉样原纤维，这些原纤维是与多种毁灭性疾病有关，包括阿尔茨海默病、帕金森病、II 型糖尿病和透析相关淀粉样变性 (DRA)，其中一种蛋白质是人类 β-2-。微球蛋白 (β2m) 在 Cu(II) 存在下可形成淀粉样原纤维，从而导致 DRA。淀粉样蛋白形成的许多一般方面已被了解，分子水平的信息淀粉样蛋白形成反应的早期阶段才刚刚开始被揭示。然而，这对于合理开发淀粉样蛋白疾病疗法至关重要。我们小组之前的工作开发并应用了新的共价标记 (CL) 技术与质谱 (MS) 一起对前淀粉样蛋白寡聚体进行结构表征从这些新工具中获得的见解有助于识别可以的小分子。抑制 β2m 淀粉样蛋白的形成了解这些抑制剂的分子基础非常重要。为了改进它们，我们将推进新的基于 MS 的方法来获得这种理解。 MS 正在成为研究蛋白质和蛋白质结构的强大工具寡聚物，但最常见的是使用基于 MS 的单独技术来收集所需的我们建议结合基于 MS 的结构工具，共同提供信息。表征 β2m 淀粉样蛋白抑制剂分子基础的协同信息。具体来说，我们将探索 CL/MS 和氢/氘交换的结合 (HDX)/MS 更详细地描述 β2m 及其结构变化我们还将建立 HDX 增强离子淌度方法来表征。在某些抑制剂存在下聚集的异构寡聚物。迄今为止，我们对 β2m 淀粉样蛋白形成的研究揭示了 Cu(II) 如何部分展开 β2m，使其在成熟原纤维产生之前形成独特的寡聚体。开始研究β2m淀粉样蛋白形成的抑制剂与基于MS的方法的结合。这里提出的旨在揭示β2m淀粉样蛋白形成抑制剂的分子基础，包括异构低聚物的表征。这项研究的一个具体成果将是更好地在分子水平上了解如何抑制 β2m 淀粉样蛋白的形成，这将导致这项工作中开发的技术将普遍适用于 DRA。其他淀粉样蛋白系统。