Visualizing Chemoreceptor Arrays in Bacterial Minicells by Cryo-Electron Tomography and Subtomogram Analysis
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.
KeywordsCryo-electron tomography Chemoreceptor array Core signaling complex Subtomogram averaging Minicell Macromolecular assembly
This work was supported in part by grants R01AI087946 from the NIAID, R01GM110243 from the NIGMS, and AU-1714 from the Welch Foundation. The direct electron detector was funded by NIH award S10OD016279.
- 12.Morado DR, Hu B, Liu J (2016) Using Tomoauto: a protocol for high-throughput automated cryo-electron tomography. J Vis Exp 107:e53608Google Scholar