Protein Self Assembly into Nanoaggregates

蛋白质自组装成纳米聚集体

基本信息

批准号：
9147607
负责人：
YURI L LYUBCHENKO
金额：
$ 38.18万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9147607
关键词：
Alzheimer&apos s Disease Amyloid Amyloid Proteins Amyloid beta-Protein Amyloidosis Atomic Force Microscopy Binding Sites Cells Computer Analysis Computer Simulation Data Deposition Development Diagnostic Disease Encephalopathies Event Failure Fluorescence Foundations Funding Goals Health Human Pathology Huntington Disease Immunoassay Individual Knowledge Lead Measures Methods Mission Modeling Molecular Morphology Nanoarray Analytical Device Nanotechnology Nature Neurodegenerative Disorders Parkinson Disease Pathology Pattern Peptides Physiological Polymers Prevention strategy Preventive Preventive Intervention Preventive treatment Process Property Proteins Public Health Raman Spectrum Analysis Research Resolution Role Spectrum Analysis Staging Structure Testing Text Therapeutic Therapeutic Intervention Toxic effect Translating Work alpha synuclein amyloid formation base dimer flexibility human disease improved innovation intermolecular interaction molecular dynamics monomer mutant nano nanoassembly neurotoxic novel novel strategies protein aggregate protein aggregation protein misfolding research study self assembly single molecule therapeutic target tool translational study

项目摘要

DESCRIPTION (provided by applicant): Aggregation of amyloid proteins is associated with a wide range of human pathologies termed protein misfolding or deposition neurodegenerative disorders, which include Alzheimer's, Parkinson's, and Huntington's diseases. It has been shown that oligomeric species of amyloid aggregates are neurotoxic. Still, the nature of these species remains unknown. Despite this importance of oligomeric species with respect to toxicity as well as in normal physiological events, knowledge regarding the molecular mechanisms underlying proteins self-assembly is very limited. The objective of this application is to characterize each oligomer at a level that will allow us to understand the molecular mechanism of the nanoassembly process. However, the fact that oligomers are formed transiently essentially impedes their characterization. To overcome this obstacle, we propose a novel approach in which oligomers of a defined size are assembled on a polymer- based Flexible Nanoarray (FNA), which will enable the use of various methods for their characterization. Based on data obtained during the current funding period, we hypothesize that the self-assembly is driven by increased, size-dependent, intermolecular interaction and stability of the oligomers. To test this hypothesis, we will thoroughly characterize FNA-assembled oligomers by applying a set of single-molecule approaches, combined with detailed computational analyses. Guided by strong preliminary data, we will text our major hypothesis through the following three specific aims: Aim 1) Develop a novel flexible nanoarray approach to measure interactions within oligomers; Aim 2) Directly measure directly the lifetimes of oligomers using a novel, tethered approach; and Aim 3) Demonstrate secondary structural analysis for individual aggregated amyloids using a Tip-Enhanced Raman Spectroscopy (TERS) approach. The rationale for the proposed aims is that understanding fundamental mechanisms of protein misfolding and aggregation has the strong potential to translate into specific approaches to control the aggregation process. These advances are expected to lead to the development of new and innovative preventative strategies and treatments for protein misfolding diseases like Alzheimer's disease. The application is innovative, because it presents a novel approach to the protein aggregation phenomenon and develops a set of new nanotechnology methods with broad biomedical applications. The proposed research is significant because the findings will lay the foundation for efficient treatments against protein misfolding diseases at the very early stages. Additionally, the availability of oligomers of select sizes assembled as FNAs opens prospects for their use as targets in the development of diagnostic tools such as immunoassays. Moreover, given that oligomers, rather than larger aggregates including fibrils, are considered neurotoxic species, the availability of oligomers with desired sizes will open realistic prospects for the development of efficient immunological preventive, diagnostic, and therapeutic strategies for Alzheimer's disease, Parkinson's disease, and other neurodegenerative disorders.

描述（由申请人提供）：淀粉样蛋白的聚集与多种被称为蛋白质错误折叠或沉积神经退行性疾病的人类病理学相关，其中包括阿尔茨海默病、帕金森病和亨廷顿病。已经表明，淀粉样蛋白聚集体的寡聚体具有神经毒性。尽管如此，这些物种的性质仍然未知，尽管寡聚物种在毒性以及正常生理事件中具有重要意义，但人们仍了解蛋白质背后的分子机制。自组装的目的是在一定程度上表征每种低聚物，使我们能够了解纳米组装过程的分子机制，但是，低聚物是瞬时形成的这一事实本质上阻碍了它们的表征。为了克服这一障碍，我们提出了一种新方法，将特定尺寸的低聚物组装在基于聚合物的柔性纳米阵列（FNA）上，这将使得能够根据当前资助期间获得的数据使用各种方法对其进行表征。在此期间，我们发现自组装是由寡聚物增加的、尺寸依赖性的、分子间相互作用和稳定性驱动的。为了检验这一假设，我们将通过应用一组单分子方法来彻底表征 FNA 组装的寡聚物。在详细的计算分析的指导下，我们将通过以下三个具体目标来阐述我们的主要假设：目标 1) 开发一种新型灵活的纳米阵列方法来测量低聚物内的相互作用；目标 2) 直接测量目标 3) 使用尖端增强拉曼光谱 (TERS) 方法对单个聚集的淀粉样蛋白进行二级结构分析。聚集具有转化为控制聚集过程的特定方法的巨大潜力，预计这些进展将导致针对蛋白质错误折叠疾病等新型创新预防策略和治疗的发展。该应用具有创新性，因为它提出了一种解决蛋白质聚集现象的新方法，并开发了一套具有广泛生物医学应用的新纳米技术方法。这项研究具有重要意义，因为这些发现将为有效治疗蛋白质错误折叠奠定基础。此外，作为 FNA 组装的选定尺寸的寡聚体的可用性为它们在开发免疫分析等诊断工具中提供了前景。此外，考虑到寡聚体，而不是更大的聚集体，包括。原纤维被认为是神经毒性物质，具有所需尺寸的寡聚物的可用性将为阿尔茨海默病、帕金森病和其他神经退行性疾病的免疫预防、诊断和治疗策略的开发开辟现实的前景。