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-Structure paper
タイトル | Subnanometer-resolution structure determination in situ by hybrid subtomogram averaging - single particle cryo-EM. |
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ジャーナル・号・ページ | Nat Commun, Vol. 11, Issue 1, Page 3709, Year 2020 |
掲載日 | 2020年7月24日 |
![]() | Ricardo M Sanchez / Yingyi Zhang / Wenbo Chen / Lea Dietrich / Mikhail Kudryashev / ![]() |
PubMed 要旨 | Cryo-electron tomography combined with subtomogram averaging (StA) has yielded high-resolution structures of macromolecules in their native context. However, high-resolution StA is not commonplace ...Cryo-electron tomography combined with subtomogram averaging (StA) has yielded high-resolution structures of macromolecules in their native context. However, high-resolution StA is not commonplace due to beam-induced sample drift, images with poor signal-to-noise ratios (SNR), challenges in CTF correction, and limited particle number. Here we address these issues by collecting tilt series with a higher electron dose at the zero-degree tilt. Particles of interest are then located within reconstructed tomograms, processed by conventional StA, and then re-extracted from the high-dose images in 2D. Single particle analysis tools are then applied to refine the 2D particle alignment and generate a reconstruction. Use of our hybrid StA (hStA) workflow improved the resolution for tobacco mosaic virus from 7.2 to 4.4 Å and for the ion channel RyR1 in crowded native membranes from 12.9 to 9.1 Å. These resolution gains make hStA a promising approach for other StA projects aimed at achieving subnanometer resolution. |
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手法 | EM (サブトモグラム平均) |
解像度 | 5.24 - 9.1 Å |
構造データ | ![]() EMDB-10834: ![]() EMDB-10840: |
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