Detection and structural analysis of misfolded proteins causing neurodegeneration

引起神经变性的错误折叠蛋白的检测和结构分析

• 批准号: 8286425
• 负责人: DAVID W COLBY
• 金额: $ 20.8万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8286425
• 关键词: Adopted; Affect; Alzheimer' s Disease; Amyloid; Amyloid fibers; Animal Model; Animals; Atomic Force Microscopy; Atrophic; Award; Binding; Biochemical; Biological Assay; Brain; Cell Culture Techniques; Cell Line; Cell model; Cells; Cerebrum; Cessation of life; Characteristics; Corpus striatum structure; Cultured Cells; Deposition; Detection; Digestion; Disease; Disease Progression; Disease model; Functional disorder; Human; Huntington Disease; In Vitro; Individual; Infection; Inherited; Laboratories; Laboratory Animal Models; Light; Mentors; Modeling; Molecular; Molecular Conformation; Mus; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Otsuka brand of aripiprazole; Parkinson Disease; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Peptide Hydrolases; Peptides; Phase; Play; Principal Investigator; Prion Diseases; Prions; Process; Property; Protein Conformation; Protein Fragment; Proteins; Recombinant Proteins; Recombinants; Research; Resistance; Role; Sampling; Seeds; Staging; Structure; System; Techniques; Thioflavin T; Toes; Work; amyloid formation; base; brain tissue; conformer; disease phenotype; human Huntingtin protein; human tissue; induced pluripotent stem cell; monomer; mouse model; polyglutamine; prion-based; programs; protein aggregate; protein misfolding; relating to nervous system; research study; stem

Aberrant folding of proteins is a feature comnnon to many neurodegenerative diseases, including prion disease, Huntington's disease (HD), Alzheimer's disease, and others. Misfolded proteins may adopt several distinct tertiary and quaternary conformations, both in terms of their stable, end states, and in terms of meta-stable intermediate states they acquire as they progress from monomers to large aggregated protein deposits in the brain. Identification of specific protein conformations involved in disease processes is lacking for many neurodegenerative diseases, although we have made progress in this regard for prion disease during the mentored phase ofthis award (Colby et.al., PNAS 2009; Colby etal., PLoS Pathog. 2010). We have begun to elucidate the manner in which biophysical properties of misfolded proteins correlate to and even directly result in specific disease phenotypes. in HD however, the role of misfolded proteins in pathogenesis is controversial and much work remains to be done to establish the conformational basis of this disease. In the independent phase of this award, we will apply the same approaches we have leamed over the past several years studying prion disease to HD in order to identify specific conformations ofthe huntingtin protein that result in neurodegeneration. Using peptides, recombinant protein, and misfolded proteins isolated from laboratory animal models and cell culture models, as well as HD patients, we will assess the extent to which the huntingtin protein is capable of acquiring diverse conformations. We will then determine if any of these conformations are capable of initiating neural death or dysfunction in cell and animal models. In a complementary approach, we will also devielop a technique for detecting misfolded huntingtin based on the propensity of misfolded proteins to nucleate or 'seed' amyloid formation. An analogous assay for detecting prions had femtogram sensitivity (Colby et. al., PNAS, 2007). The ability to sensitively detect misfolded huntingtin will enable us to identify its presence much earlier in disease progression than is currently possible. A better understanding ofthe molecular basis of Huntington's disease and related neurodegenerative disorders shall aid substantially in the search for a cure.

蛋白质的异常折叠是许多神经退行性疾病的特征，包括prion 疾病，亨廷顿病（HD），阿尔茨海默氏病等。错误折叠的蛋白质可能会采用几种 从稳定，最终状态和 当它们从单体发展到大型蛋白质时，它们获得的元稳定中间状态 沉积在大脑中。鉴定疾病过程中涉及的特定蛋白质构象的IS 尽管我们在这方面取得了进展，但缺乏许多神经退行性疾病 在这一奖项的指导阶段期间的疾病（Colby等，PNAS，2009； Colby等，PLOS病原体。 2010）。我们已经开始阐明错误折叠蛋白的生物物理特性的方式 与甚至直接导致特定疾病表型相关。然而，在高清中，错误折叠的作用 发病机理中的蛋白质是有争议的，要建立构象仍有许多工作要做 这种疾病的基础。在该奖项的独立阶段，我们将采用与我们采用的相同方法 在过去的几年中，研究prion疾病的高清以识别特定构象 导致神经退行性的亨廷汀蛋白。使用肽，重组蛋白和 从实验室动物模型和细胞培养模型以及HD患者中分离出的错误折叠的蛋白质， 我们将评估亨廷汀蛋白能够获得多种构象的程度。我们 然后将确定这些构型是否能够在细胞中引发神经死亡或功能障碍 和动物模型。在一种互补的方法中，我们还将Devielop一种用于检测错误折叠的技术 基于错误折叠蛋白对成核或“种子”淀粉样蛋白形成的倾向。一个 用于检测王室的类似测定具有依据图（Colby et al。，PNAS，2007）。能力 敏感地检测错误折叠的亨廷顿将使我们能够在疾病中更早地识别其存在 进展比目前可能的进展。更好地理解亨廷顿的分子基础 疾病和相关的神经退行性疾病应大大有助于寻找治愈方法。