Development of novel crosslinking-mass spectrometry technologies to interrogate the molecular sociology of cells

研究背景生物过程由瞬间相互作用的蛋白质组合组成的复杂网络进行。揭示细胞的这种“分子社会学”对于理解细胞内稳态和疾病机制是至关重要的，但要实现这一点，迫切需要新的工具。目的和目标-目标是：化学交联和质谱学相结合可以用来识别细胞中蛋白质-蛋白质的相互作用，以揭示这一“分子社会学”。然而，需要新的交联剂来克服实验挑战。在本项目中，您将设计/合成新型化学交联剂，并验证这些试剂用于细胞中的蛋白质结构/相互作用分析。潜在的应用和益处这个跨学科的项目将涵盖合成和分析化学、生物化学和细胞生物学。您将设计和合成一组交联剂，这些交联剂具有解决细胞结构生物学的交联化工作流程中遇到的方法学挑战的功能。接下来，您将验证这些交联剂设计的分析工作流程，以便通过基于质谱学的蛋白质组学进行体外和细胞内蛋白质相互作用分析。最后，您将通过研究心血管疾病和蛋白质错误折叠/聚集疾病(例如帕金森氏症和阿尔茨海默病)的细胞模型来举例说明这些新型交联剂作为化学工具的功能。其结果将是一系列具有优化性能的交联剂，将改变我们直接在细胞中询问生物机制的能力。研究领域：细胞中的生物机制。有待获得的资格：博士学位

Context of researchBiological processes are undertaken by an intricate network of transiently interacting protein assemblies. Uncovering this 'molecular sociology' of the cell is vital to understanding cellular homeostasis and disease mechanisms, but to achieve this, new tools are urgently needed. Aims and objectives - The objectives are:Chemical crosslinking combined with mass spectrometry can be used to identify protein-protein interactions in cells, to reveal this 'molecular sociology'. However, new crosslinkers are needed to overcome experimental challenges. In this project you will design/synthesise novel chemical crosslinkers and validate these reagents for protein structure/interaction analysis in cells. Potential applications and benefitsThis interdisciplinary project will encompass synthetic and analytical chemistry, biological chemistry and cell biology. You will design and synthesize a panel of crosslinkers with functionalities that address the methodological challenges encountered in crosslinking workflows for cellular structural biology. Next, you will validate analytical workflows for these crosslinker designs for in vitro and in cell protein interaction analysis by mass spectrometry-based proteomics. Finally, you will exemplify the functionality of these novel crosslinkers as chemical tools by studying cellular models of cardiovascular disease and protein misfolding/aggregation diseases (e.g. Parkinson's and Alzheimer's diseases). The outcome will be a range of crosslinking reagents with optimised properties that will transform our ability to interrogate biological mechanisms directly in cells.Research Areas: Biological mechanisms in cells.Qualification to be attained: PhD degree