Amyloid Transcription Block Survival - Deriving Functionally Active Peptide Inhibitors of Amyloidosis and Toxicity

淀粉样蛋白转录阻断存活 - 衍生淀粉样变性和毒性的功能活性肽抑制剂

• 批准号: 2598875
• 金额: --
• 依托单位: University of Bath
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2598875
• 关键词: Amyloid; Transcription; Block; Survival; Deriving

The aim of this project is to develop peptides that can be used to effectively inhibit formation of a toxic protein responsible for the pathology of Parkinson's disease (PD). The protein, known as alpha-synuclein (aS), self-associates inside dopamine producing cells in the brain to form toxic clumps known as Lewy bodies that interfere with normal brain function and lead to the symptoms of the disease (Meade et al, Mol Neurodegen 2019: https://rdcu.be/bLohx; Meade et al NPJ Parkinson's disease 2020). The PhD program will focus the student on inhibiting this process building from a system that we have already demonstrated to work (Cheruvara et al, J.Biol.Chem 2015). Using a novel screening system that targets aS prior to any misfolding or aggregation, the student will screen large peptide libraries (>2 Million members) inside living bacterial cells using a 'Transcription Block Survival' (TBS) assay. In this assay, inhibitors will only become selected if DNA-binding activity is prevented, leading to a restoration of cell viability. Our overarching aim is to assign function to specific sequence elements within our newly generated inhibitors to demonstrate the principles of rational inhibitor design, ultimately improving the properties of future peptide generations. To achieve this the student will use the TBS assay to generate numerous inhibitory peptides to block the very first steps in the misfolding of aS. This will provide a wide range of sequences from which we can understand the mechanism of inhibition using biophysical, neuronal cell-based methods. The project will also involve work with Bristol University and the application of high-resolution NMR to probe structures and interactions of the peptides with aS, providing valuable structural biology expertise. Finally, by comparing endogenously produced peptides against externally added peptides, the student will begin to explore aspects of drug delivery, such as permeability and the ability to reach intracellular targets. The composition of the supervisory team ensures comprehensive expertise in all facets of this interdisciplinary project. You will find a supportive and stimulating training environment, to guide you through the challenges and rewards of this project while gaining a wide range of skills that are translatable to many other systems.

该项目的目的是开发可用于有效抑制导致帕金森病（PD）病理学的毒性蛋白质形成的肽。这种蛋白质被称为α-突触核蛋白（aS），在大脑中的多巴胺产生细胞内自我缔合，形成称为路易体的有毒团块，干扰正常的大脑功能并导致疾病的症状（米德等人，Mol Neurodegen 2019：https：//rdcu.be/bLohx;米德等人NPJ帕金森病2020）。博士课程将重点关注学生从我们已经证明有效的系统中抑制该过程的构建（Cheruvara et al，J.Biol.Chem 2015）。使用一种新的筛选系统，在任何错误折叠或聚集之前靶向aS，学生将使用“转录块生存”（TBS）测定筛选活细菌细胞内的大型肽库（> 200万成员）。在该试验中，只有当DNA结合活性被阻止时，抑制剂才会被选择，从而导致细胞活力的恢复。我们的首要目标是将功能分配给我们新生成的抑制剂中的特定序列元件，以证明合理抑制剂设计的原则，最终改善未来肽代的特性。为了实现这一目标，学生将使用TBS测定来产生许多抑制肽，以阻断aS错误折叠的最初步骤。这将提供一个广泛的序列，我们可以从中了解使用生物物理，神经元细胞为基础的方法的抑制机制。该项目还将涉及与布里斯托大学的合作，以及高分辨率NMR的应用，以探测肽与aS的结构和相互作用，提供宝贵的结构生物学专业知识。最后，通过比较内源性产生的肽与外部添加的肽，学生将开始探索药物输送的各个方面，如渗透性和到达细胞内靶点的能力。监督团队的组成确保了这个跨学科项目各个方面的全面专业知识。你会发现一个支持和激励的培训环境，引导你通过这个项目的挑战和奖励，同时获得广泛的技能，可翻译到许多其他系统。