Department of Microbiology and Molecular Genetics, McGovern Medical School, Houston, TX, USA.
Methods Mol Biol. 2024;2727:17-25. doi: 10.1007/978-1-0716-3491-2_2.
The cell division machinery or "divisome" of many bacteria, including Escherichia coli, contains homologs of tubulin (FtsZ) and actin (FtsA) that interact with each other to promote the synthesis of septal peptidoglycan. FtsA oligomers have an essential role as a track for tethering dynamically treadmilling FtsZ protofilaments to the cytoplasmic membrane. Other bacterial cytoskeletal oligomers such as MreB also assemble on and move along the membrane. Structures of these oligomers on membranes in vitro may mimic their behavior in the cell. Here, we describe a protocol to visualize FtsA oligomeric structures on membranes and their interactions with FtsZ protofilaments using negative stain transmission electron microscopy along with tomography.
许多细菌（包括大肠杆菌）的细胞分裂机制或“分裂体”含有微管蛋白（FtsZ）和肌动蛋白（FtsA）的同源物，它们相互作用促进隔膜肽聚糖的合成。FtsA 寡聚体作为一个轨道，对于将动态行走的 FtsZ 原纤维附着到细胞质膜上起着重要作用。其他细菌细胞骨架寡聚体，如 MreB，也在膜上组装并沿膜移动。在体外膜上这些寡聚体的结构可能模拟它们在细胞中的行为。在这里，我们描述了一种使用负染色透射电子显微镜和断层摄影术可视化膜上 FtsA 寡聚体结构及其与 FtsZ 原纤维相互作用的方案。
