Unravelling the structure and conformational dynamics of membrane proteins using H/D-exchange mass spectrometry and cryo-EM

使用 H/D 交换质谱和冷冻电镜揭示膜蛋白的结构和构象动力学

• 批准号: 2593790
• 金额: --
• 依托单位: University of Southampton
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2593790
• 关键词: Unravelling; structure; conformational; dynamics; membrane

Membrane proteins are biological macromolecules that are present within dynamic and complex cellular membranes. They have diverse complex cellular functions and represent more than half of drug targets; however, the molecular mechanisms governing their modes of action remain poorly understood. Most membrane proteins are highly dynamic and undergo structural and conformational rearrangements to perform their function. Whilst Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS) and Cryo-Electron Microscopy (cryo-EM) each provide unique information, both techniques complement and validate each other providing static pictures as well as dynamics and flexibility information of large and complex systems1. This proposal focuses on developing workflows to understand the structure and conformational dynamics of membrane proteins using HDX-MS and cryo-EM. This project will focus on developing the knowledge and expertise of the PhD student in both HDX-MS and cryo-EM by studying the efflux pump (MdtF). The first HDX-MS and native MS measurements of membrane proteins (including AcrB) in SMALPs2,3 have been published as well as a recent publication demonstrating the impact of a clinically relevant efflux pump mutation in AcrB using HDX-MS4.

膜蛋白是存在于动态和复杂的细胞膜内的生物大分子。它们具有多种复杂的细胞功能，占药物靶点的一半以上;然而，对其作用模式的分子机制仍知之甚少。大多数膜蛋白是高度动态的，并经历结构和构象重排来执行其功能。虽然氢/氘交换质谱（HDX-MS）和低温电子显微镜（cryo-EM）各自提供独特的信息，但这两种技术相互补充和验证，提供大型复杂系统的静态图片以及动态和灵活性信息1。该提案的重点是开发工作流程，以了解使用HDX-MS和cryo-EM的膜蛋白的结构和构象动力学。该项目将通过研究外排泵（MdtF），重点发展博士生在HDX-MS和cryo-EM方面的知识和专业知识。已发表SMALP中膜蛋白（包括AcrB）的首次HDX-MS和天然MS测量结果2，3，以及最近的一篇出版物，证明了使用HDX-MS的AcrB中临床相关外排泵突变的影响4。