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一种导致帕金森氏病(PD)病理的有毒蛋白的形成的多肽。这种蛋白质被称为α-突触核蛋白(AS)，它在大脑中的多巴胺产生细胞内自我结合，形成被称为路易小体的有毒团块，干扰正常的大脑功能，并导致疾病的症状(Meade等人，MOL Neurodegen 2019年：https://rdcu.be/bLohx；Meade等人，2020年)。博士课程将把学生的重点放在从我们已经证明有效的系统中抑制这一过程的建立(Cheruvara等人，J.Biol.Chem 2015)。使用一种新的筛选系统，目标是在任何错误折叠或聚集之前，学生将使用‘转录阻断生存’(TBS)分析在活的细菌细胞中筛选大型多肽库(&gt；200万成员)。在这项试验中，只有在阻止DNA结合活性，导致细胞活力恢复的情况下，才会选择抑制剂。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素，以演示合理的抑制剂设计原则，最终改善未来多肽的性质。为了实现这一点，学生将使用TBS分析来产生许多抑制肽来阻止AS错误折叠的第一步。这将提供广泛的序列，从中我们可以了解抑制的机制使用生物物理，神经细胞为基础的方法。该项目还将涉及与布里斯托尔大学的合作和高分辨率核磁共振的应用，以探测多肽的结构和与AS的相互作用，提供宝贵的结构生物学专业知识。最后，通过比较内部产生的多肽和外部添加的多肽，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。监督小组的组成确保了这一跨学科项目所有方面的全面专业知识。你会发现一个支持性和激励性的培训环境，指导你通过这个项目的挑战和回报，同时获得广泛的技能，这些技能可以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.