Structural Characterization of Amyloid Fibrils Using Deep UV Raman Spectroscopy

使用深紫外拉曼光谱法表征淀粉样原纤维的结构

• 批准号: 8447473
• 负责人: IGOR K LEDNEV
• 金额: $ 19.21万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-15 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8447473
• 关键词: Adopted; Alzheimer' s Disease; Amyloid; Amyloid Fibrils; Amyloidosis; Appearance; Complement; Creutzfeldt-Jakob Syndrome; Deuterium; Diagnostic radiologic examination; Disease; Employee Strikes; Familial Amyloid Neuropathies; Future; Genetic Polymorphism; Goals; HIV; Heterogeneity; Huntington Disease; Hydrogen; Hydrogen Bonding; In Vitro; Isotope Labeling; Kinetics; Knowledge; Label; Laboratories; Length; Literature; Methodology; Methods; Modeling; Molecular Conformation; Morphology; Nature; Neurodegenerative Disorders; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Parkinson Disease; Patients; Peptide Fragments; Peptides; Phosphoric Monoester Hydrolases; Prion Diseases; Prions; Process; Property; Prostatic; Protein Precursors; Proteins; Raman Spectrum Analysis; Reaction; Reporting; Roentgen Rays; Role; Seminal fluid; Sexual Transmission; Solutions; Structural Models; Structure; Symptoms; System; Testing; Therapeutic; Time; Toxic effect; Variant; Vertebral column; X-Ray Crystallography; amyloid fibril formation; base; comparative; design; improved; in vivo; molecular dynamics; neutrophil; novel; particle; polypeptide; protein aggregate; protein misfolding; public health relevance; small molecule; solid state; solid state nuclear magnetic resonance; structural biology; therapeutic development; tool; ultraviolet

DESCRIPTION (provided by applicant): Amyloid fibrils are associated with numerous debilitating diseases such as Alzheimer's disease, Parkinson's disease, Huntington's diseases, prion disease, familial amyloid polyneuropathy, senile systemic amyloidosis and type II diabetes, etc. The rational design of successful therapeutic strategies calls for detailed structural characterization of amyloid fibrils. Solid-state NMR and X-ray microcrystallography applied to short peptide fragments of amyloidogenic proteins have contributed greatly to our knowledge about the fibril core structure. The main goal of this project is to extend our understanding of the amyloid fibril structure by utilizing deep ultraviolet resonance Raman spectroscopy combined with hydrogen-deuterium exchange. This new method, recently pioneered in our laboratory, does not require isotope labeling and creates the opportunity for bridging the gap between our extended knowledge in the structural variations of model short peptide microcrystals and the core structure of amyloid fibrils prepared from entire proteins. We will determine and classify the core structure of amyloid fibrils prepared from full-length proteins. It has been postulated that differences in infectivity of prion protein aggregates, and related differences in the apparent symptoms of Creutzfeldt-Jakob disease patients, are possible due to amyloid polymorphism. Small molecules could potentially manipulate aggregate morphology and provide the basis for therapeutic approaches to suppress the formation of toxic aggregates. Therefore, it is critical to (i) have a robust method for structural characterization of polymorphs, (ii) understand the nature of amyloid polymorphs and their toxicity, and (iii) eventually, be able to control (suppress) the formation of toxic polymorphs. We will test alternative hypothesis concerning the level of structure at which functional polymorphisms are realized. When fully developed, in future studies, the proposed approach will make possible the comparative structural characterization of in vivo and in vitro amyloid fibrils. This will bridge the gap between in vivo and in vitro studies of neurodegenerative diseases. In particular, the method will provide unique information revealing the structural determinants of infectious and non-infectious amyloids.

描述(申请人提供)：淀粉样纤维与许多衰弱的疾病有关，如阿尔茨海默病、帕金森氏病、亨廷顿病、Pron病、家族性淀粉样多发性神经病、老年性系统性淀粉样变性和II型糖尿病等。成功的治疗策略的合理设计要求淀粉样纤维具有详细的结构特征。固体核磁共振和X射线微晶学应用于淀粉样蛋白的短肽片段，对我们了解纤维核心结构有很大的贡献。这个项目的主要目标是通过利用深紫外共振拉曼光谱结合氢-氚交换来扩大我们对淀粉样原纤维结构的理解。这种新方法是我们实验室最近首创的，不需要同位素标记，并为弥合我们在模型短肽微晶体结构变化方面的扩展知识与由整个蛋白质制备的淀粉样纤维的核心结构之间的差距创造了机会。我们将对由全长蛋白质制备的淀粉样纤维的核心结构进行测定和分类。据推测，可能是由于淀粉样蛋白的多态，导致普恩蛋白聚合体的感染性差异，以及克雅氏病患者明显症状的相关差异。小分子可能会潜在地操纵聚集体的形态，并为抑制有毒聚集体形成的治疗方法提供基础。因此，关键是(I)有一种可靠的方法来表征多态的结构，(Ii)了解淀粉样多型的性质和它们的毒性，以及(Iii)最终能够控制(抑制)有毒多态的形成。我们将测试关于实现功能多态的结构水平的替代假设。当完全发展，在未来的研究中，所提出的方法将使体内和体外淀粉样纤维的比较结构特征成为可能。这将弥合神经退行性疾病的体内和体外研究之间的差距。特别是，该方法将提供独特的信息，揭示传染性和非传染性淀粉样蛋白的结构决定因素。