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  Subnanometer-resolution structure determination in situ by hybrid subtomogram averaging - single particle cryo-EM

Sanchez, R. M., Zhang, Y., Chen, W., Dietrich, L., & Kudryashev, M. (2020). Subnanometer-resolution structure determination in situ by hybrid subtomogram averaging - single particle cryo-EM. Nature Communications, 11: 3709. doi:10.1038/s41467-020-17466-0.

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 Creators:
Sanchez, Ricardo M.1, 2, Author           
Zhang, Yingyi1, 2, Author           
Chen, Wenbo1, 2, Author           
Dietrich, Lea3, Author           
Kudryashev, Misha1, 2, Author           
Affiliations:
1Sofja Kovalevskaja Group, Max Planck Institute of Biophysics, Max Planck Society, ou_2253651              
2Buchmann Institute for Molecular Life Sciences, Goethe-University of Frankfurt, 60348 Frankfurt am Main, Germany, ou_persistent22              
3Department of Structural Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068291              

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 Abstract: 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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Language(s): eng - English
 Dates: 2020-05-072020-06-302020-07-24
 Publication Status: Published online
 Pages: 7
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1038/s41467-020-17466-0
 Degree: -

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Title: Nature Communications
  Abbreviation : Nat. Commun.
Source Genre: Journal
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Publ. Info: London : Nature Publishing Group
Pages: - Volume / Issue: 11 Sequence Number: 3709 Start / End Page: - Identifier: ISSN: 2041-1723
CoNE: https://pure.mpg.de/cone/journals/resource/2041-1723