Vadas O, Jenkins M L, Dornan G L, Burke J E
Pharmaceutical Sciences Section, University of Geneva, Geneva, Switzerland.
University of Victoria, Victoria BC, Canada.
Methods Enzymol. 2017;583:143-172. doi: 10.1016/bs.mie.2016.09.008. Epub 2016 Oct 18.
Many fundamental cellular processes are controlled via assembly of a network of proteins at membrane surfaces. The proper recruitment of proteins to membranes can be controlled by a wide variety of mechanisms, including protein lipidation, protein-protein interactions, posttranslational modifications, and binding to specific lipid species present in membranes. There are, however, only a limited number of analytical techniques that can study the assembly of protein-membrane complexes at the molecular level. A relatively new addition to the set of techniques available to study these protein-membrane systems is the use of hydrogen-deuterium exchange mass spectrometry (HDX-MS). HDX-MS experiments measure protein conformational dynamics in their native state, based on the rate of exchange of amide hydrogens with solvent. This review discusses the use of HDX-MS as a tool to identify the interfaces of proteins with membranes and membrane-associated proteins, as well as define conformational changes elicited by membrane recruitment. Specific examples will focus on the use of HDX-MS to examine how large macromolecular protein complexes are recruited and activated on membranes, and how both posttranslational modifications and cancer-linked oncogenic mutations affect these processes.
许多基本的细胞过程是通过蛋白质网络在膜表面的组装来控制的。蛋白质向膜的正确募集可通过多种机制来控制，包括蛋白质脂化、蛋白质-蛋白质相互作用、翻译后修饰以及与膜中存在的特定脂质种类结合。然而，只有有限数量的分析技术能够在分子水平上研究蛋白质-膜复合物的组装。用于研究这些蛋白质-膜系统的一系列技术中，相对较新的一项是氢-氘交换质谱（HDX-MS）的使用。HDX-MS实验基于酰胺氢与溶剂的交换速率来测量蛋白质在其天然状态下的构象动力学。本综述讨论了使用HDX-MS作为一种工具来识别蛋白质与膜及膜相关蛋白的界面，以及定义由膜募集引发的构象变化。具体例子将聚焦于使用HDX-MS来研究大型高分子蛋白质复合物如何在膜上被募集和激活，以及翻译后修饰和与癌症相关的致癌突变如何影响这些过程。
