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

该项目的目的是开发可用于有效抑制帕金森病（PD）病理负责的有毒蛋白形成的肽。这种被称为α -突触核蛋白（as）的蛋白质在大脑中产生多巴胺的细胞内自我结合，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症状（Meade等人，Mol Neurodegen 2019: https://rdcu.be/bLohx； Meade等人NPJ帕金森病2020）。博士课程将重点关注学生如何从我们已经证明有效的系统中抑制这一过程（Cheruvara等人，j.b oll . 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，该学生将使用“转录阻断存活”（TBS）试验筛选活细菌细胞内的大型肽库（bbb20万个成员）。在这个实验中，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。我们的首要目标是在我们新生成的抑制剂中为特定序列元素分配功能，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实验生成许多抑制肽来阻止aS错误折叠的第一步。这将提供广泛的序列，从中我们可以使用生物物理，基于神经元细胞的方法来理解抑制机制。该项目还将涉及与布里斯托尔大学的合作，并应用高分辨率核磁共振来探测肽与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多肽和外部添加的多肽，学生将开始探索药物传递的各个方面，如渗透性和到达细胞内目标的能力。监督团队的组成确保了在这个跨学科项目的各个方面都有全面的专业知识。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回报，同时获得广泛的技能，可翻译到许多其他系统。

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.