Native ambient mass spectrometry for membrane proteins

膜蛋白的天然环境质谱分析

• 批准号: EP/Y004604/1
• 负责人: Helen Cooper
• 金额: $ 68.89万
• 依托单位: University of Birmingham
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FY004604%2F1
• 关键词: Native; ambient; mass; spectrometry; membrane

Proteins are large biomolecules that perform all the functions required for life. The myriad roles and uses of proteins are not simply down to their chemical architecture but also the shapes into which they fold. The study of proteins therefore requires a tool capable of characterization at the molecular level, which provides information on both chemical and overall three-dimensional structure. Mass spectrometry is such a tool, and has provided startling insights into the mechanisms by which proteins operate.This proposal relates to a class of proteins known as membrane proteins. Membrane proteins are entangled with other molecules that make up the cell membrane, indeed need those molecules to maintain their shape. They present a conduit from the outside of a cell to its inner workings, and consequently represent ~ 70% of drug targets for conditions from high blood pressure and heart disease, to stroke and cancer. Membrane proteins can also be hijacked by viruses, facilitating their entry into the cell and again providing a focal point for the development of therapeutics. Despite their potential therapeutic value, structural information on membrane proteins lags behind that available for more accessible proteins because of the challenges of solubilising and maintaining the shape of the protein during analysis. We have developed native ambient mass spectrometry (NAMS), a tool for molecular analysis of proteins and their interacting partners directly from tissue. NAMS both provides structural information and enables their spatial distribution to be visualised. To date, NAMS has been primarily applied to accessible, soluble proteins. Preliminary data, however, suggest that the approach is also suitable for the imaging and analysis of membrane proteins. The aim of this proposal is to develop NAMS specifically for membrane proteins. We will design, build and validate a source for NAMS which enables targeted delivery of reagents to facilitate extraction of membrane proteins from tissue. Two approaches for extraction will be investigated: 1) use of solubilising detergents and 2) capture of membrane proteins together with some of the surrounding membrane molecules in a "styrene maleic acid lipid particle" or SMALP. Alternative polymers to styrene maleic acid will also be considered.

蛋白质是大型生物分子，可执行生命所需的所有功能。蛋白质的无数作用和用途不仅归结于它们的化学结构，还归结为它们折叠的形状。因此，对蛋白质的研究需要一种能够在分子水平表征的工具，该工具提供了有关化学和总体三维结构的信息。质谱是这样的工具，并且为蛋白质操作的机制提供了令人震惊的见解。该建议与一类称为膜蛋白的蛋白质有关。膜蛋白与构成细胞膜的其他分子纠缠在一起，确实需要这些分子来保持其形状。他们呈现从细胞外部到其内部起作用的导管，因此，约有70％的药物靶标的高血压和心脏病到中风和癌症的疾病。膜蛋白也可以被病毒劫持，促进其进入细胞，并再次为疗法开发提供了焦点。尽管具有潜在的治疗价值，但有关膜蛋白的结构信息落在后面的蛋白质后面，因为在分析过程中溶解和维持蛋白质的形状的挑战。我们已经开发了天然环境质谱法（NAM），这是一种直接从组织中直接从组织中对蛋白质及其相互作用伴侣进行分子分析的工具。 NAM都提供结构信息，并使它们的空间分布可视化。迄今为止，NAM已主要应用于可访问的可溶性蛋白。但是，初步数据表明该方法也适用于膜蛋白的成像和分析。该建议的目的是专门为膜蛋白开发NAM。我们将设计，构建和验证NAM的来源，该源可以靶向递送试剂以促进从组织中提取膜蛋白。将研究两种提取方法：1）使用溶解洗涤剂和2）捕获膜蛋白以及“苯乙烯马来酸脂质颗粒”或SMALP中的一些周围膜分子。还将考虑到苯乙烯马来酸的替代聚合物。