TY  - JOUR
A1  - Sánchez Loayza, Ricardo Miguel
A1  - Zhang, Yingyi
A1  - Chen, Wenbo
A1  - Dietrich, Lea
A1  - Kudryashev, Mikhail
T1  - Subnanometer-resolution structure determination in situ by hybrid subtomogram averaging - single particle cryo-EM
T2  - Nature Communications
N2  - 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.
Y1  - 2020
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/79897
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-798977
SN  - 2041-1723
VL  - 11
IS  - 3709
PB  - Nature Publishing Group UK
CY  - London
ER  -