Structural biology/ biochemistry--alpha synuclein & other PD linked gene products

结构生物学/生物化学--α突触核蛋白

• 批准号: 6302892
• 负责人: PETER T LANSBURY
• 金额: $ 29.46万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6302892
• 关键词: Lewy body; Parkinson's disease; alpha synuclein; atomic force microscopy; combinatorial chemistry; disease /disorder onset; enzyme structure; esterase; gene mutation; genetic disorder; molecular assembly /self assembly; polymerization; protein biosynthesis; protein degradation; protein folding; protein sequence; protein structure function; recombinant proteins; site directed mutagenesis; structural biology; ubiquitin

The overall goal of these experiments is to determine the effects of PD-linked mutations on the properties of the gene products and to use this information to discover methods to test possible explanations for pathogenicity. We expect that this work will generate new therapeutic strategies for the treatment of Parkinson's disease (PD). Our emphasis will be on protein fibrillogenesis, since fibrillar cytoplasmic aggregates, or Lewy bodies, are diagnostic for PD and a major fibrillar component of Lewy bodies is also the product of a gene linked to early-onset PD. Three mutations, in two different genes, encoding alpha-synuclein (alphaS) and ubiquitin C-hydrolase (UCH), have been linked to early-onset PD. We have shown that the two alphaS mutations effect the oligomerization properties of the protein; both favor oligomerization. It is a central goal of the proposed research to understand the structural basis for oligomerization and fibrillization and the relationship between this process and disease (the latter will require a collaboration between this project and project 3). We are also very interested in the ubiquitin-dependent degradation of alphaS, especially since UCH may be involved in that pathway. Finally, the possibility that mutant UCH may also be a fibrillogenic protein is under investigation. Protein (UCH and alphaS) fibrillization will be a target for medium-throughput screening assays to be run in the Center core facility (Core B). A gene linked to juvenile-onset parkinsonism, parkin, will be the subject of future biochemical and biophysical investigations. This protein contains an N-terminal ubiquitin homology domain, which suggests its involvement (like UCH) in the degradative process. We intend to characterize wild-type and mutant forms of Parkin. The fact that this disease is inherited in an autosomal recessive manner suggests that gain of function due to toxic oligomers may not be involved.

这些实验的总体目标是确定pd相关突变对基因产物特性的影响，并利用这些信息发现测试致病性可能解释的方法。我们期望这项工作将为帕金森病（PD）的治疗提供新的治疗策略。我们的重点将是蛋白质纤维的形成，因为纤维胞质聚集体或路易小体是PD的诊断，路易小体的主要纤维成分也是与早发性PD相关的基因的产物。编码α -突触核蛋白（α has）和泛素c -水解酶（UCH）的两个不同基因的三种突变与早发性帕金森病有关。我们已经证明，两个α - has突变影响蛋白质的寡聚化特性；两者都有利于寡聚化。拟议研究的中心目标是了解寡聚化和成纤维化的结构基础以及这一过程与疾病之间的关系（后者将需要本项目与项目3之间的合作）。我们也对α - has的泛素依赖性降解非常感兴趣，特别是因为UCH可能参与了这一途径。最后，突变的UCH也可能是一种纤维原蛋白的可能性正在研究中。蛋白（UCH和alpha has）纤溶化将是在中心核心设施（core B）中进行中通量筛选分析的目标。一种与青少年发病的帕金森氏症有关的基因，帕金森氏症，将是未来生物化学和生物物理学研究的主题。该蛋白含有一个n端泛素同源结构域，这表明它参与了降解过程（如UCH）。我们打算鉴定野生型和突变型帕金病。这种疾病以常染色体隐性方式遗传的事实表明，由于毒性低聚物而获得的功能可能与此无关。