2006Wang Pitch

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Citation

Wang YA, Yu X, Yip C, Strynadka NC, Egelman EH (2006) Structural polymorphism in bacterial EspA Wlaments revealed by cryo-EM and an improved approach to helical reconstruction. Structure 14:1189–1196

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Abstract

The traditional Fourier-Bessel approach to three-dimensional reconstruction from electron microscopic (EM) images of helical polymers involves averaging over filaments, assuming a homogeneous structure and symmetry. We have used a real-space reconstruction approach to study the EspA filaments formed by enteropathogenic E. coli. In negative stain, the symmetry of these filaments is ambiguous, and we suggest that such ambiguities may be more prevalent than realized. Using cryo-EM of frozen-hydrated filaments, we find that these filaments have a fixed twist with 5.6 subunits per turn but an axial rise per subunit that varies from about 3.6 A to 5.6 A. Reconstructions at approximately 15 A resolution show a switching between the more compressed and extended filaments in the packing of putative alpha helices around the hollow lumen. Outside of a crystal, where there is nothing to maintain long-range order, the structural polymorphism in helical polymers may be much greater than has been assumed.

Keywords

Pitch correction helical structures

Links

Article http://www.ncbi.nlm.nih.gov/pubmed/16843900

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