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蛋白通过组织癌前细胞的自我毁灭来预防癌症。因此，p53在大多数癌症中都出现了故障，并允许不受控制的细胞生长，这是该疾病的核心。因此，了解p53功能背后的分子机制是至关重要的，研究人员现在知道p53在一定程度上是由大量的翻译后修饰（PTMs）调节的。在这个项目中，我将研究非典型PTM对p53的功能影响：通过顺式和反式立体异构体之间的脯氨酸异构化（Pro-Iso）来重新定位蛋白质的骨架。具体来说，我假设Pro-Iso作为一个“构象开关”，驱使p53进入不同的功能状态。由于难以获得化学上明确定义的p53变异，以及p53对结构生物学技术的抗拒，这一假设尚未得到验证。为了克服这些挑战，我将通过跨学科的化学生物学和生物物理学方法探索Pro-Iso在p53功能中的作用。首先，我将使用我的导师Manuel m<e:1> ller博士（伦敦国王学院）开创的蛋白质半合成技术来生产全长p53变体，这些变体带有精确的化学修饰，可以探测Pro-Iso的不同方面。其次，我将对这些变体进行一系列生化分析，以确定关键特性和功能的差异。最后，我将使用单分子Förster共振能谱来研究Pro-Iso对p53分子相互作用下构象动力学的影响。这个项目将为我提供作为一名独立研究小组负责人所需的可转移技能、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，从而扩大知识库，全球利益相关者将从中寻求解决老龄化世界中不断增长的癌症发病率。