Optimizing and defining the mechanism of alpha-synuclein disaggregation by Hsp104

优化和定义 Hsp104 α-突触核蛋白解聚机制

• 批准号: 8316617
• 负责人: Morgan DeSantis
• 金额: $ 4.22万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8316617
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Affect; Age of Onset; Alzheimer' s Disease; Amyloid; Amyloid fibers; Binding; Bradykinesia; Brain; Calorimetry; Characteristics; Collaborations; Couples; Data; Deterioration; Disease; Dyes; Effectiveness; Electron Microscopy; Ensure; Fiber; Goals; Hydrolysis; In Vitro; Individual; Lewy Bodies; Link; Mediating; Missense Mutation; Modeling; Molecular Conformation; Monitor; Movement Disorders; Mutation; Nature; Nerve Degeneration; Neurodegenerative Disorders; North America; Nucleotides; Parkinson Disease; Patients; Peptides; Positioning Attribute; Protein Conformation; Proteins; Rattus; Resistance; Role; Sedimentation process; Severities; Shapes; Structure; Substantia nigra structure; Symptoms; System; Testing; Therapeutic; Therapeutic Agents; Titrations; Toxic effect; Tremor; Yeasts; alpha synuclein; amyloid formation; amyloid structure; base; brain tissue; dopaminergic neuron; effective therapy; mutant; pars compacta; protein aggregate; protein aggregation; protein misfolding; protein structure; research study; stoichiometry; synuclein; therapeutic target; translocase; yeast protein

DESCRIPTION (provided by applicant): Misfolding and aggregation of the protein ?-synuclein (? -syn) is associated with Parkinson's Disease (PD), which is a devastating neurodegenerative movement disorder. In PD, ? -syn misfolds into an aberrant protein conformation, termed amyloid, which is highly stable, possesses a characteristic cross-?? structure, and is largely viewed to be an intractable condition. There is evidence that pre- amyloid oligomers of ? -syn possess greater toxicity than the fibers themselves. Furthermore, certain rare mutations of ? -syn are associated with familial forms of PD and often result in a decreased age of onset and increased severity of symptoms. These mutants have been shown to increase propensity of ? -syn aggregation or to increase the concentration of the pre-amyloid oligomers. A remarkable yeast protein, Hsp104, has been shown to ameliorate symptoms of ? -syn aggregation in a rat model of PD by disaggregating the ? -syn fibers and oligomers. However, high concentrations of Hsp104 were required to observe disaggregation, which reduces the effectiveness of this potential PD therapeutic. Additionally, it is not clear how Hsp104 is able to remodel ? -syn fibers and oligomers, which obscures ways in which this protein may be further developed to treat PD. We have isolated an Hsp104 mutant with even greater potency towards ? -syn fibers, which gives us confidence that Hsp104 will be eventually become a viable PD treatment. The goal of this proposal is to optimize Hsp104 for even higher anti-?? -syn fiber and oligomer activity. We plan on targeting both wild-type and disease associated mutants of ? -syn to ensure that Hsp104 can be used to treat all cases of PD. We also aim to understand the mechanism of Hsp104 disaggregation of ? -syn fibers and oligomers in order to facilitate even further optimization of Hsp104 as a PD therapeutic. PUBLIC HEALTH RELEVANCE: Parkinson's Disease is a devastating neurodegenerative disorder which is characterized by protein misfolding and aggregation in brain tissue. These misfolded proteins, termed amyloid, are very stable and difficult to clear. However, the yeast protein, Hsp104 is able to reverse protein misfolding and clear the amyloid aggregates. We are focusing on two things: First, we will optimize Hsp104 activity so it is more potent against the amyloid aggregates found in Parkinson's patients; and second, we will determine how Hsp104 functions.

描述（由申请人提供）：蛋白质β-突触核蛋白（β-syn）的错误折叠和聚集与帕金森病（PD）有关，帕金森病是一种破坏性的神经退行性运动障碍。在PD中，？ -syn 错误折叠成一种异常的蛋白质构象，称为淀粉样蛋白，它高度稳定，具有特征性的交叉？结构，并且在很大程度上被认为是一个棘手的问题。有证据表明，前淀粉样蛋白寡聚体？ -syn比纤维本身具有更大的毒性。此外，某些罕见的突变？ -syn 与家族性 PD 相关，通常会导致发病年龄降低和症状严重程度增加。这些突变体已被证明可以增加 ? 的倾向。 -syn 聚集或增加前淀粉样蛋白寡聚体的浓度。一种非凡的酵母蛋白 Hsp104 已被证明可以改善 ? -syn 在 PD 大鼠模型中通过分解 ? 进行聚合-syn纤维和低聚物。然而，需要高浓度的 Hsp104 才能观察到解聚，这降低了这种潜在 PD 治疗方法的有效性。另外，目前还不清楚Hsp104是如何重塑的？ -syn纤维 和寡聚物，这掩盖了进一步开发这种蛋白质来治疗帕金森病的方法。我们已经分离出一种 Hsp104 突变体，其对 ? -syn 纤维，这让我们相信 Hsp104 最终将成为一种可行的 PD 治疗方法。该提案的目标是优化 Hsp104 以获得更高的抗 ?? -syn纤维和低聚物活性。我们计划针对野生型和疾病相关突变体？ -syn 确保 Hsp104 可用于治疗所有 PD 病例。我们还旨在了解 Hsp104 分解的机制？ -syn 纤维和寡聚物，以促进 Hsp104 作为 PD 治疗剂的进一步优化。 公共健康相关性：帕金森病是一种破坏性的神经退行性疾病，其特征是脑组织中蛋白质错误折叠和聚集。这些错误折叠的蛋白质被称为淀粉样蛋白，非常稳定且难以清除。然而，酵母蛋白 Hsp104 能够逆转蛋白质错误折叠并清除淀粉样蛋白聚集物。我们专注于两件事：首先，我们将优化 Hsp104 活性，使其更有效地对抗帕金森病患者中发现的淀粉样蛋白聚集体；其次，我们将确定 Hsp104 的功能。