Abstract
Using a liquid-helium-cooled superconducting electron cryo-microscope, we obtained low-dose images of negatively stained preparations at 4 K and collected structural data to 1/9.6 Å-1 for flagellar filaments from the strain SJW117 of Salmonella typhimurium (serotype gt). The subunits of this left-handed, straight filament are non-helically perturbed in a pairwise manner. The perturbation corresponds to an alternating conformation in every other row of subunits. These are the 5-start rows and, necessarily, the resulting structure has a seam. The perturbation is not confined to the outside but extends into the structure. We separated the non-symmetric and symmetric parts of the structural data and generated a three-dimensional reconstruction from the latter. The resulting density map is a structure similar in domain organization to the left-handed filament of S. typhimurium SJW1660. Filtered images generated from the non-symmetric component show an ordered and polar structure. The nature of the perturbation was analyzed by model building using a sphere to represent the subunit at low resolution. A lateral shift of ~10°mimics the perturbation.
| Original language | English |
|---|---|
| Pages (from-to) | 759-773 |
| Number of pages | 15 |
| Journal | Journal of Molecular Biology |
| Volume | 276 |
| Issue number | 4 |
| DOIs | |
| State | Published - 6 Mar 1998 |
Keywords
- Bacterial flagellar filaments
- Cryo-electron microscopy
- Helical perturbations
- Negative staining
- Three-dimensional helical reconstruction
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