Qin Zhuan, Hu Bo, Liu Jun
Department of Microbial Pathogenesis and Microbial Sciences Institute, Yale School of Medicine, Advanced Biosciences Center, New Haven, CT, USA.
Department of Microbiology and Molecular Biology, McGovern Medical School, The University of Texas, Houston, TX, USA.
Methods Mol Biol. 2018;1729:187-199. doi: 10.1007/978-1-4939-7577-8_17.
Bacterial chemoreceptors form a highly ordered array in concert with the CheA kinase and the CheW coupling protein. The precise architecture of the array is responsible for high sensitivity, high dynamic range, and strong amplification of chemotaxis signaling. Cryo-electron tomography (cryo-ET) has emerged as a unique tool to visualize bacterial chemotaxis arrays at molecular level. Here we describe a detailed cryo-ET and subtomogram averaging procedure to determine in situ structure of the chemoreceptor arrays in Salmonella minicells. The procedure should be readily applicable to visualize other large macromolecular assemblies in their cellular context.
细菌化学感受器与CheA激酶和CheW偶联蛋白协同形成高度有序的阵列。该阵列精确的结构负责化学感受信号的高灵敏度、高动态范围和强放大作用。冷冻电子断层扫描（cryo-ET）已成为在分子水平上可视化细菌化学感受阵列的独特工具。在这里，我们描述了一种详细的冷冻电子断层扫描和亚断层平均程序，以确定鼠伤寒沙门氏菌小细胞中化学感受器阵列的原位结构。该程序应易于应用于在细胞环境中可视化其他大型大分子组装体。
