p53PRISM: Regulation of life-and-death decisions by conformational switches

p53PRISM：构象开关调节生死决定

• 批准号: EP/Z000823/1
• 负责人: Manuel Muller
• 金额: $ 24.5万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FZ000823%2F1
• 关键词: p53PRISM; Regulation; life; death; decisions

The protein p53 guards against cancer by orchestrating the self-destruction of precancerous cells. Accordingly, p53 in most cancers ismalfunctioning and permits the unchecked cellular growth central to the disease. Understanding the molecular mechanismsunderlying p53's function is thus crucial, and researchers now know that p53 is regulated in part by a plethora of post-translationalmodifications (PTMs).In this project, I will investigate the functional consequences in p53 of an atypical PTM: the reorientation of a protein's backbone bythe isomerisation of a proline between cis and trans stereoisomers (Pro-Iso). Specifically, I hypothesise Pro-Iso acts as a"conformational switch" which drives p53 into distinct functional states. This hypothesis is untested due to the difficulty of obtainingchemically well-defined p53 variants and p53's recalcitrance towards structural biology techniques.To overcome these challenges, I will explore the role of Pro-Iso in p53's function via an interdisciplinary chemical biology andbiophysical approach. First, I will use the protein semisynthesis techniques pioneered by my supervisor Dr. Manuel Müller (King'sCollege London) to produce full-length p53 variants bearing precise chemical modifications that probe different aspects of Pro-Iso.Second, I will subject these variants to a host of biochemical assays to ascertain differences in key properties and functions. Finally, Iwill use single-molecule Förster resonance energy spectroscopy to investigate the effect of Pro-Iso on the conformational dynamicsunderlying p53's molecular interactions.This project will provide me with the transferable skills, training and track record needed to pursue a career as an independentresearch group leader. More importantly, this research will help further decode how p53 makes cellular life-and-death decisions andthus expand the knowledgebase from which global stakeholders seek to tackle growing cancer rates in an aging world.

p53 蛋白通过协调癌前细胞的自我毁灭来预防癌症，因此，p53 在大多数癌症中都存在功能障碍，并允许不受控制的细胞生长成为疾病的核心，因此了解 p53 功能的分子机制至关重要。 p53 部分受到大量翻译后修饰 (PTM) 的调节。在这个项目中，我将研究 p53 中的功能后果非典型 PTM：通过顺式和反式立体异构体 (Pro-Iso) 之间的脯氨酸异构化来重新定位蛋白质骨架。具体来说，我假设 Pro-Iso 充当“构象开关”，驱动 p53 进入不同的功能状态。由于难以获得化学上明确的 p53 变体以及 p53 对结构生物学技术的抵制，因此未经测试。为了克服这些挑战，我将探索其作用首先，我将使用我的导师 Manuel Müller 博士（伦敦国王学院）首创的蛋白质半合成技术来生产带有精确化学修饰的全长 p53 变体。探究 Pro-Iso 的不同方面。其次，我将对这些变体进行一系列生化测定，以确定关键特性和功能的差异。最后，我将使用单分子。 Förster 共振能谱研究 Pro-Iso 对 p53 分子相互作用的构象动力学的影响。该项目将为我提供作为独立研究小组领导者所需的可转移技能、培训和跟踪记录，更重要的是，这项研究。将有助于进一步解码p53如何做出细胞生死决定，从而扩大全球利益相关者寻求解决老龄化世界中日益增长的癌症发病率的知识库。