Department of Life Sciences, Imperial College London, London, UK.
Mol Microbiol. 2021 Mar;115(3):366-382. doi: 10.1111/mmi.14637. Epub 2020 Dec 5.
"CryoEM" has come of age, enabling considerable structural insights into many facets of molecular biology. Here, we present a primer for microbiologists to understand the capabilities and limitations of two complementary cryoEM techniques for studying bacterial secretion systems. The first, single particle analysis, determines the structures of purified protein complexes to resolutions sufficient for molecular modeling, while the second, electron cryotomography and subtomogram averaging, tends to determine more modest resolution structures of protein complexes in intact cells. We illustrate these abilities with examples of insights provided into how secretion systems work by cryoEM, with a focus on type III secretion systems.
"CryoEM" 已经成熟,能够为分子生物学的许多方面提供重要的结构见解。在这里,我们为微生物学家提供了一个入门指南,以了解两种互补的 cryoEM 技术用于研究细菌分泌系统的能力和局限性。第一种是单颗粒分析,它可以确定纯化蛋白复合物的结构,分辨率足以进行分子建模,而第二种是电子冷冻断层扫描和子断层平均法,通常可以确定完整细胞中蛋白复合物更适中的分辨率结构。我们用 cryoEM 提供的关于分泌系统如何工作的见解示例来说明这些能力,重点是 III 型分泌系统。