Cryo-electron tomography (cryo-ET) is a powerful tool for generating images of three-dimensional structures of biological specimens. However, it is limited to imaging thin structures. The study of membrane-protein complexes in intact bacterial membranes by cryo-ET remains challenging owing to the thickness of typical bacteria. To bypass this problem, Fu et al. generated bacterial 'ghosts' for use in cryo-ET by expressing the phage FX174 lysis gene E in Escherichia coli. Gene E expression leads to spot lysis, which allows the cytoplasm to escape while preserving the cell membrane and cell shape. By carrying out cryo-ET on these ghosts, the researchers were able to generate a high-resolution tomographic reconstruction of chemotaxis receptor signaling complexes within the cell membrane. This method holds promise for facilitating structural analysis of membrane-protein complexes in a native context.
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Bacterial ghosts for cryo-electron tomography. Nat Methods 12, 105 (2015). https://doi.org/10.1038/nmeth.3266
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DOI: https://doi.org/10.1038/nmeth.3266