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: 8054840
• 负责人: IGOR K LEDNEV
• 金额: $ 20.33万
• 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/8054840
• 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. PUBLIC HEALTH RELEVANCE: Successful outcomes of the proposed study will not only improve our understanding of amyloid fibril structure in general and the nature of amyloid polymorphs and their toxicity in particular, but also provide the basis for the development of therapeutic approaches to suppress the formation of toxic aggregates. When fully developed, the proposed approach will make possible the comparative structural characterization of in vivo and in vitro amyloid fibrils and bridge the gap between in vivo and in vitro studies of neurodegenerative diseases.

说明（申请人提供）：淀粉样蛋白原纤维与许多衰弱性疾病有关，如阿尔茨海默病、帕金森病、亨廷顿病、朊病毒病、家族性淀粉样蛋白多发性神经病、老年系统性淀粉样变性和II型糖尿病等。成功治疗策略的合理设计需要淀粉样蛋白原纤维的详细结构表征。固态核磁共振和X-射线显微晶体学应用于短肽片段的淀粉样蛋白质有很大的贡献，我们的知识的原纤维核心结构。本计画的主要目的是利用深紫外共振拉曼光谱结合氢氘交换技术，来扩展我们对淀粉样蛋白原纤维结构的了解。这种新的方法，最近在我们的实验室开创，不需要同位素标记，并创造了弥合我们的扩展知识之间的差距差距模型短肽微晶的结构变化和从整个蛋白质制备的淀粉样蛋白原纤维的核心结构的机会。我们将确定和分类的核心结构的淀粉样纤维制备全长蛋白质。据推测，朊病毒蛋白聚集体的感染性差异，以及克雅氏病患者的明显症状的相关差异，可能是由于淀粉样蛋白多态性。小分子可以潜在地操纵聚集体形态，并为抑制毒性聚集体形成的治疗方法提供基础。因此，至关重要的是（i）具有用于多晶型物的结构表征的稳健方法，（ii）了解淀粉样蛋白多晶型物的性质及其毒性，以及（iii）最终能够控制（抑制）有毒多晶型物的形成。我们将检验关于实现功能多态性的结构水平的备择假设。当充分发展，在未来的研究中，所提出的方法将有可能在体内和体外淀粉样纤维的比较结构表征。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，揭示感染性和非感染性淀粉样蛋白的结构决定因素。 公共卫生相关性：这项研究的成功结果不仅将提高我们对淀粉样蛋白原纤维结构的理解，特别是淀粉样蛋白多晶型物的性质及其毒性，而且还为开发抑制毒性聚集体形成的治疗方法提供了基础。当充分发展，所提出的方法将有可能在体内和体外淀粉样纤维的比较结构表征和桥梁之间的差距差距在体内和体外研究的神经退行性疾病。