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Visualizing Chemoreceptor Arrays in Bacterial Minicells by Cryo-Electron Tomography and Subtomogram Analysis

  • Zhuan Qin
  • Bo Hu
  • Jun Liu
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1729)

Abstract

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.

Keywords

Cryo-electron tomography Chemoreceptor array Core signaling complex Subtomogram averaging Minicell Macromolecular assembly 

Notes

Acknowledgment

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.

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Department of Microbial Pathogenesis and Microbial Sciences InstituteYale School of Medicine, Advanced Biosciences CenterNew HavenUSA
  2. 2.Department of Microbiology and Molecular BiologyMcGovern Medical School, The University of TexasHoustonUSA

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