Cryo-EM as a tool to study bacterial efflux systems and the membrane proteome.
Cryo-EM as a tool to study bacterial efflux systems and the membrane proteome.
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Cryo-EM作为研究细菌外排系统和膜蛋白质组的工具。
DOI：
10.12703/r/10-24
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Yu EW
中科院分区：
文献类型：
--
作者：
Klenotic PA;Morgan CE;Yu EW
抗生素耐药性是对全球健康的新威胁。目前对这些类型的细菌感染的治疗方案变得越来越不充分。因此，需要新的创新技术来帮助识别和表征新药和药物靶标，这对于对抗多重耐药菌株至关重要。细菌外排系统已经成为药物设计的一个有吸引力的目标，因为阻断它们的输出功能显著增加了所施用抗生素的效力。然而，为了开发有效的和可耐受的外排泵抑制剂具有高的功效，详细的结构信息是必需的载脂蛋白和底物结合形式的这些膜蛋白。低温电子显微镜（cryo-EM）的出现极大地推进了膜蛋白结构生物学领域。它显著增强了解决大型多蛋白复合物的能力，以及从异质样品中提取有意义的数据的能力，例如从单个数据集中识别细菌核糖体的几种组装状态。这种技术可以扩展到解决基板结合的外排泵和整个外排系统从以前无法使用的膜蛋白样品制备的结构。随后，cryo-EM与其他生物物理技术相结合，有可能显着推进膜蛋白结构生物学领域。识别完整的转运机制，酶信号转导途径和其他膜相关复合物的能力将有助于我们充分理解膜蛋白质组的复杂性。
Antibiotic resistance is an emerging threat to global health. Current treatment regimens for these types of bacterial infections are becoming increasingly inadequate. Thus, new innovative technologies are needed to help identify and characterize novel drugs and drug targets which are critical in order to combat multidrug-resistant bacterial strains. Bacterial efflux systems have emerged as an attractive target for drug design, as blocking their export function significantly increases the potency of administered antibiotics. However, in order to develop potent and tolerable efflux pump inhibitors with high efficacy, detailed structural information is required for both the apo- and substrate-bound forms of these membrane proteins. The emergence of cryo-electron microscopy (cryo-EM) has greatly advanced the field of membrane protein structural biology. It has significantly enhanced the ability to solve large multi-protein complexes as well as extract meaningful data from a heterogeneous sample, such as identification of several assembly states of the bacterial ribosome, from a single data set. This technique can be expanded to solve the structures of substrate-bound efflux pumps and entire efflux systems from previously unusable membrane protein sample preparations. Subsequently, cryo-EM combined with other biophysical techniques has the potential to markedly advance the field of membrane protein structural biology. The ability to discern complete transport machineries, enzymatic signal transduction pathways, and other membrane-associated complexes will help us fully understand the complexities of the membrane proteome.