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- W2913382246 abstract "Tubular structures are common occurrences in biological systems. Tubules can be formed from lipid membranes that have been deformed by a protein lattice or only ordered protein oligomerization as is the case of filaments. In many cases there is substantial heterogeneity in the widths of the tubules, the proteins that make up the tubules, or proteins that decorate the tubules. This heterogeneity can create a challenge when attempting to use cryo-EM to determine the structures of the tubular proteins. The common method of using helical symmetry to resolve the structure will average out areas of heterogeneity, removing areas of interesting. We have developed a method called Reconstruction of Average Subtracted Tubular Regions (RASTR) to deal with heterogeneity by treating the tubules not as helical filaments and relying on symmetry, but instead focusing on a segment of one surface of the tubule. During RASTR, we first align the tubules vertically, then determine the azimuthal global average of the tubules. This is then reconstructed in 3D which gives a smoothed-out representation of the surface of the tubules. A segment of the smooth 3D tubule is masked out leaving a tubule with a missing segment on one surface. This is then projected and subtracted from individual cryo-EM images of the tubules, resulting in images of the tubule with everything but one surface strongly down-weighted. The up-weighted surface can then be treated as a single particle and can be aligned and classified by single particle methods. A benefit of our method is that it limits the degrees of freedom of alignment, which can allow us to drop particles that align outside a narrow range. Here we will present our results comparing the reconstruction of generated model data and real data of the same tubules (VIP tubules)." @default.
- W2913382246 created "2019-02-21" @default.
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- W2913382246 date "2019-02-01" @default.
- W2913382246 modified "2023-09-30" @default.
- W2913382246 title "Reconstruction of Average Subtracted Tubular Regions (RASTR)" @default.
- W2913382246 doi "https://doi.org/10.1016/j.bpj.2018.11.3099" @default.
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