Mapping membrane protein dynamics in time and space with mass spectrometry

利用质谱绘制膜蛋白的时间和空间动态图

• 批准号: EP/V011715/1
• 负责人: Argyris Politis
• 金额: $ 129.78万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FV011715%2F1
• 关键词: Mapping; membrane; protein; dynamics; time

The advancement of new analytical tools and methods are at the centre of tackling the biggest challenges in the life sciences. One such unmet challenge is understanding the structural dynamics underpinning function in membrane-embedded proteins. Membrane proteins control what comes in and what goes out of the cell. As a consequence, they constitute the main targets of more than half of known drugs. Despite their critical importance, existing methods often fail to uncover structural information about this important class of biomolecules, thus precluding progresses related to therapeutic intervention and drug discovery. More importantly, we currently lack the tools to capture the dynamics of membrane proteins within their native environment wherein they move and operate. This is primarily due to the complexity of such systems as they are embedded into a heterogeneous layer of lipids, which protect their hydrophobic core of membrane proteins. New tools are therefore urgently needed to unveil the dynamic motions of membrane proteins and allow mechanistic insights important for addressing current and future challenges related to human health and disease.Here, we will built a new method to capture molecular movies of membrane proteins in action. To do this, we will develop time-resolved hydrogen deuterium exchange mass spectrometry (tHDX-MS). HDX-MS is a sensitive analytical tool that can accurately monitor the exchange of hydrogen atoms with the heavier deuterium in solution, thus offering information about protein dynamics. By combining the emerging HDX-MS technology with microfluidic techniques, we will enable snapshots of membrane protein states in times ranging from microseconds to hours and with adjustable resolution. To enable applicability of our strategy within the native lipid environment wherein membrane proteins function, we will utilise the controlled patches of membrane biomimetics, the so called nanodiscs. The nanodisc technology will allow us to fine-tune the lipid composition surrounding membrane proteins and assess the individual effect of specific lipids on membrane protein structure and dynamics. We will demonstrate applicability of our approach on a range of important systems of increasing size and complexity including the challenging G protein-coupled receptors (GPCRs) that are the key drug targets. To make our approach amenable to large and dynamic complexes and circumvent current challenges with respect to sensitivity and resolution, we will work with our industrial partner (Waters Corp.) to utilise a currently non-commercial, prototypical instrumentation (Cyclic HDX-MS). This together with our methodological advancements will allow us to be the first to achieve this for membrane proteins in lipid context and thus become the leaders in this rapidly evolving field of research in the UK and worldwide. Overall, this fellowship will not only establish a new tool for tackling key challenges in deciphering the dynamic mechanisms underpinning membrane protein function but it will also allow me to lead this exciting and emerging field of research, currently under-represented in the UK.

新的分析工具和方法的进步是应对生命科学中最大的挑战的中心。这样的挑战的一个是理解膜包裹蛋白中基础功能的结构动力学。膜蛋白控制进来的东西以及从细胞中出现的东西。结果，它们构成了一半以上已知药物的主要靶标。尽管它们的重要性至关重要，但现有方法通常无法揭示有关这类重要的生物分子的结构信息，因此排除了与治疗干预和药物发现有关的进展。更重要的是，我们目前缺乏在其本地环境中捕获膜蛋白动态的工具，在其本地环境中它们移动和运行。这主要是由于这些系统的复杂性嵌入到异质的脂质层中，从而保护其膜蛋白的疏水核心。因此，迫切需要新的工具来揭示膜蛋白的动态动作，并允许机械洞察力对于解决与人类健康和疾病有关的当前和未来挑战很重要的机械见解。在这里，我们将建立一种新方法来捕捉作用中膜蛋白的分子电影。为此，我们将开发时间分辨的氢氘交换质谱法（THDX-MS）。 HDX-MS是一种敏感的分析工具，可以准确监测溶液中氢原子的交换，从而提供有关蛋白质动力学的信息。通过将新兴的HDX-MS技术与微流体技术相结合，我们将在从微秒到小时以及可调节分辨率的时间内启用膜蛋白态的快照。为了使我们的策略适用于膜蛋白功能的天然脂质环境中，我们将利用所谓的纳米尺度的膜仿生片的受控斑块。纳米盘技术将使我们能够微调围绕膜蛋白的脂质组成，并评估特定脂质对膜蛋白结构和动力学的个体影响。我们将证明我们的方法适用于大小和复杂性的一系列重要系统，包括具有挑战性的G蛋白偶联受体（GPCR）是关键药物靶标。为了使我们的方法适应大型动态的复合物，并规避当前的敏感性和分辨率挑战，我们将与我们的工业合作伙伴（Waters Corp.）合作，利用当前非商业的原型仪器（环状HDX-MS）。这与我们的方法论进步将使我们成为在脂质环境中第一个实现膜蛋白的人，因此成为英国和全球迅速发展的研究领域的领导者。总体而言，该研究金不仅将建立一个新工具来应对膜蛋白功能支撑的动态机制时要面临的关键挑战，而且还可以使我能够领导这个令人兴奋和新兴的研究领域，目前在英国代表性不足。

项目成果

Chromatographic phospholipid trapping for automated H/D exchange mass spectrometry analysis of membrane protein-lipid assemblies

色谱磷脂捕获用于膜蛋白-脂质组件的自动 H/D 交换质谱分析

• DOI: 10.26434/chemrxiv-2022-8j01g
• 发表时间: 2022
• 作者: Hammerschmid D

A neutralizing epitope on the SD1 domain of SARS-CoV-2 spike targeted following infection and vaccination.

• DOI: 10.1016/j.celrep.2022.111276
• 发表时间: 2022-08-23
• 期刊: CELL REPORTS
• 影响因子: 8.8
• 作者: Seow, Jeffrey;Khan, Hataf;Rosa, Annachiara;Calvares, Valeria;Graham, Carl;Pickering, Suzanne;Pye, Valerie E.;Cronin, Nora B.;Huettner, Isabella;Malim, Michael H.;Politis, Argyris;Cherepanov, Peter;Doores, Katie J.
• 通讯作者: Doores, Katie J.

Structural dynamics in the evolution of SARS-CoV-2 spike glycoprotein

SARS-CoV-2 刺突糖蛋白进化的结构动力学

• DOI: 10.21203/rs.3.rs-2049401/v1
• 发表时间: 2022
• 作者: Calvaresi V

Cyclic ion mobility for hydrogen/deuterium exchange-mass spectrometry applications

• DOI: 10.26434/chemrxiv-2023-gzfhv
• 发表时间: 2023-12-20
• 期刊: chemRxiv
• 作者: Griffiths，Damon;Anderson，Malcolm;Politis，Argyris
• 通讯作者: Politis，Argyris

Chromatographic Phospholipid Trapping for Automated H/D Exchange Mass Spectrometry of Membrane Protein-Lipid Assemblies.

• DOI: 10.1021/acs.analchem.2c04876
• 发表时间: 2023-02-07
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Hammerschmid, Dietmar;Calvaresi, Valeria;Bailey, Chloe;Lewis, Benjamin Russell;Politis, Argyris;Morris, Michael;Denbigh, Laetitia;Anderson, Malcolm;Reading, Eamonn
• 通讯作者: Reading, Eamonn