Department of Chemistry, King's College London, Britannia House, 7 Trinity Street, London SE1 1DB, U.K.
Essays Biochem. 2023 Mar 29;67(2):255-267. doi: 10.1042/EBC20220190.
Multidrug efflux pumps are ubiquitous across both eukaryotes and prokaryotes, and have major implications in antimicrobial and multidrug resistance. They reside within cellular membranes and have proven difficult to study owing to their hydrophobic character and relationship with their compositionally complex lipid environment. Advances in structural mass spectrometry (MS) techniques have made it possible to study these systems to elucidate critical information on their structure-function relationships. For example, MS techniques can report on protein structural dynamics, stoichiometry, connectivity, solvent accessibility, and binding interactions with ligands, lipids, and other proteins. This information proving powerful when used in conjunction with complementary structural biology methods and molecular dynamics (MD) simulations. In the present review, aimed at those not experts in MS techniques, we report on the current uses of MS in studying multidrug efflux systems, practical considerations to consider, and the future direction of the field. In the first section, we highlight the importance of studying multidrug efflux proteins, and introduce a range of different MS techniques and explain what information they yield. In the second section, we review recent studies that have utilised MS techniques to study and characterise a range of different multidrug efflux systems.
多药外排泵在真核生物和原核生物中普遍存在，对抗菌和多药耐药性有重大影响。它们位于细胞膜内，由于其疏水性和与组成复杂的脂质环境的关系，研究起来颇具难度。结构质谱 (MS) 技术的进步使得研究这些系统成为可能，从而阐明其结构-功能关系的关键信息。例如，MS 技术可以报告蛋白质结构的动态性、化学计量学、连通性、溶剂可及性以及与配体、脂质和其他蛋白质的结合相互作用。当与互补的结构生物学方法和分子动力学 (MD) 模拟结合使用时，这些信息非常有用。在本综述中，针对非 MS 技术专家，我们报告了 MS 在研究多药外排系统中的当前用途、需要考虑的实际问题以及该领域的未来发展方向。在第一节中，我们强调了研究多药外排蛋白的重要性，介绍了一系列不同的 MS 技术，并解释了它们提供的信息。在第二节中，我们回顾了最近利用 MS 技术研究和表征一系列不同的多药外排系统的研究。
