Faculty of Biology, Medicine and Health, School of Biological Sciences, Division of Molecular & Cellular Function, The University of Manchester, Manchester M13 9PT, U.K.
Manchester Institute of Biotechnology, University of Manchester, Princess Street, Manchester M1 7DN, U.K.
Essays Biochem. 2023 Mar 29;67(2):187-200. doi: 10.1042/EBC20220173.
Integral membrane proteins (IMPs) perform a range of diverse functions and their dysfunction underlies numerous pathological conditions. Consequently, IMPs constitute most drug targets, and the elucidation of their mechanism of action has become an intense field of research. Historically, IMP studies have relied on their extraction from membranes using detergents, which have the potential to perturbate their structure and dynamics. To circumnavigate this issue, an array of membrane mimetics has been developed that aim to reconstitute IMPs into native-like lipid environments that more accurately represent the biological membrane. Hydrogen/deuterium exchange-mass spectrometry (HDX-MS) has emerged as a versatile tool for probing protein dynamics in solution. The continued development of HDX-MS methodology has allowed practitioners to investigate IMPs using increasingly native-like membrane mimetics, and even pushing the study of IMPs into the in vivo cellular environment. Consequently, HDX-MS has come of age and is playing an ever-increasingly important role in the IMP structural biologist toolkit. In the present mini-review, we discuss the evolution of membrane mimetics in the HDX-MS context, focusing on seminal publications and recent innovations that have led to this point. We also discuss state-of-the-art methodological and instrumental advancements that are likely to play a significant role in the generation of high-quality HDX-MS data of IMPs in the future.
整合膜蛋白 (IMPs) 具有多种不同的功能，其功能障碍是许多病理状况的基础。因此，IMPs 构成了大多数药物靶点，阐明其作用机制已成为一个研究热点。从历史上看，IMP 的研究依赖于使用去污剂从膜中提取，这有可能破坏其结构和动力学。为了解决这个问题，已经开发了一系列的膜类似物，旨在将 IMP 重新构成类似于天然的脂质环境，更准确地模拟生物膜。氢/氘交换-质谱（HDX-MS）已成为探测溶液中蛋白质动力学的一种多功能工具。HDX-MS 方法的不断发展使得研究人员能够使用越来越接近天然的膜类似物来研究 IMP，甚至将 IMP 的研究推向体内细胞环境。因此，HDX-MS 已经成熟，并在 IMP 结构生物学家工具包中发挥着越来越重要的作用。在本综述中，我们讨论了 HDX-MS 背景下膜类似物的发展，重点介绍了导致这一发展的开创性出版物和最新创新。我们还讨论了最先进的方法学和仪器学进展，这些进展很可能在未来产生高质量的 IMPs HDX-MS 数据方面发挥重要作用。
