Pathological Changes of Alpha-Synuclein Structure in the Brain

好了！ α-突触核蛋白聚集体(α-Syn)是帕金森病(PD)、痴呆的病理标志 伴有路易小体(DLB)和多系统萎缩(MSA)。此外，α同步的遗传增加 α基因的表达和点突变是一种越来越多的文献记载的早期- 发病家族性帕金森病(FPD)。病理性低聚物已被广泛研究，但只有详细的分析。 来自重组来源的合成聚集体的数量是可用的。因此，我们希望将 我们从PD/DLB和MSA脑中分离到病理αSyn种。我们计划在这里进行一次广泛的 从人类患者脑中分离出的有毒和自我允许的αSyn聚集态的特征。 隔离物种的特征将是功能(毒性)和结构上的 结构-功能关系分析。对不同大脑的额外分析将允许进行比较 不同疾病(疾病“株”)之间的结构特异性。隔离程序将集中于 针对3个不同的亚细胞部位(胞浆、膜- 关联的、不可溶的)。我们的新概念的结果可能指向理性药物的令人兴奋的可能性 根据致病菌株的确切结构细节进行设计。病原菌的检测和量化 α-Syn寡聚体可作为诊断联体核病的生物标志物，并将提供新的 为社区提供试剂以及有价值的治疗平台(菌株特异性PMCA检测) 化合物筛选。为了推进我们的假设，我们建议从两个主要方面收集数据 方向：目标1：寻找人脑中是否存在天然可溶的α-syn“病态种子” PD、DLB和MSA患者非变性条件下的匀浆及其生物活性测定 在病原聚集和毒性的动态分析中。目标2：描述独特的结构 从每个疾病亚组(PD/DLB/MSA)分离出的脑不溶聚集体的特征(“菌株”) 以及它们的扩增产物，以对比它们的生化和生物物理特性与 疾病。来自我们的新概念的结果可能指向合理的PD药物设计中令人兴奋的可能性 基于致病菌株的确切结构细节。此外，检测和量化 致病性α-Syn寡聚体可作为诊断共核病的生物标记物，并将提供 新试剂(合成人脑衍生菌株)与有价值的平台一起进入社区 (菌株特异性检测试验)用于治疗化合物筛选。 好了！ 好了！

! Aggregates of alpha-synuclein (αSyn) are a pathological hallmark of Parkinson's disease (PD), Dementia with Lewy Bodies (DLB) and Multiple System Atrophy (MSA). Moreover, genetic increases in αSyn expression and point mutations in the αSyn gene are an increasingly well-documented precipitant of early- onset familial PD (fPD). Pathological oligomers have been extensively studied, but only detailed analyses of synthetic aggregates from recombinant sources are available. Therefore, we wish to characterize pathological αSyn species we are isolating from PD/DLB and MSA brains. We plan here an extensive characterization of toxic and self-permissive aggregated forms of αSyn, isolated from human patient brain. Characterization of the isolated species will be both functional (toxicity) and structural for a correlative analysis of structure-function relationship. Additional analysis of different brains will allow a comparison between structure specificity of the different diseases (disease “strains”). The isolation procedure will focus on gentle, non-denaturing methods targeting 3 different sub-cellular locales (cytosol, membrane- associated, insoluble). Results from our novel concept could point to exciting possibilities in rational drug design based on exact structural details of pathogenic strains. Detection and quantification the pathogenic αSyn oligomers may be used as a biomarker for diagnosis of synucleinopathies and will provide novel reagents to the community along with valuable platforms (strain specific PMCA assays) for therapeutic compound screening. To move our hypotheses forward, we propose to gather data in two major directions: Aim 1: Search for the existence of natively soluble `pathological seeds' of αSyn in human brain homogenates of PD, DLB and MSA patients under non-denaturing conditions and quantify their bioactivity in dynamic assays of pathogenic aggregation and toxicity. Aim 2: Characterize the unique structural features (“strains”) of the brain isolated insoluble aggregates from each disease subgroup (PD/DLB/MSA) and their amplification products to contrast their biochemical and biophysical properties in relation to disease. Results from our novel concept could point to exciting possibilities in rational PD drug design based on exact structural details of pathogenic strains. In addition, detection and quantification the pathogenic αSyn oligomers may be used as a biomarker for diagnosis of synucleinopathies and will provide novel reagents (synthetic human brain derived strains) to the community along with valuable platforms (strain specific detection assays) for therapeutic compound screening. ! !