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: 6499886
• 负责人: PETER T LANSBURY
• 金额: $ 24.81万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6499886
• 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，并利用这些信息来发现测试致病性可能解释的方法。 我们期望这项工作将为治疗帕金森病（PD）产生新的治疗策略。 我们的重点是蛋白质纤维形成，因为纤维状细胞质聚集体或路易体可以诊断帕金森病，而路易体的主要纤维成分也是与早发性帕金森病相关的基因的产物。 编码 α-突触核蛋白 (alphaS) 和泛素 C-水解酶 (UCH) 的两个不同基因中的三个突变与早发性帕金森病有关。 我们已经证明，两个 alphaS 突变会影响蛋白质的寡聚特性；两者都有利于低聚。 拟议研究的中心目标是了解寡聚化和原纤维化的结构基础以及该过程与疾病之间的关系（后者需要本项目和项目 3 之间的合作）。 我们对 alphaS 的泛素依赖性降解也非常感兴趣，特别是因为 UCH 可能参与该途径。 最后，突变 UCH 也可能是一种纤维蛋白的可能性正在研究中。 蛋白质（UCH 和 alphaS）原纤维化将成为中心核心设施（核心 B）运行的中通量筛选测定的目标。与青少年帕金森症相关的基因parkin将成为未来生化和生物物理研究的主题。 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）。 我们打算表征 Parkin 的野生型和突变型。 该疾病以常染色体隐性方式遗传的事实表明，可能不涉及有毒寡聚体导致的功能获得。