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High-resolution cryo-EM reconstructions in the presence of substantial aberrations

Here, an analysis is performed of how uncorrected antisymmetric aberrations, such as coma and trefoil, affect cryo-EM single-particle reconstruction (SPR) results, and an analytical formula quantifying information loss owing to their presence is inferred that explains why Fourier-shell coefficient-b...

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Detalles Bibliográficos
Autores principales: Bromberg, Raquel, Guo, Yirui, Borek, Dominika, Otwinowski, Zbyszek
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7201289/
https://www.ncbi.nlm.nih.gov/pubmed/32431828
http://dx.doi.org/10.1107/S2052252520002444
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author Bromberg, Raquel
Guo, Yirui
Borek, Dominika
Otwinowski, Zbyszek
author_facet Bromberg, Raquel
Guo, Yirui
Borek, Dominika
Otwinowski, Zbyszek
author_sort Bromberg, Raquel
collection PubMed
description Here, an analysis is performed of how uncorrected antisymmetric aberrations, such as coma and trefoil, affect cryo-EM single-particle reconstruction (SPR) results, and an analytical formula quantifying information loss owing to their presence is inferred that explains why Fourier-shell coefficient-based statistics may report significantly overestimated resolution if these aberrations are not fully corrected. The analysis is validated with reference-based aberration refinement for two cryo-EM SPR data sets acquired with a 200 kV microscope in the presence of coma exceeding 40 µm, and 2.3 and 2.7 Å reconstructions for 144 and 173 kDa particles, respectively, were obtained. The results provide a description of an efficient approach for assessing information loss in cryo-EM SPR data acquired in the presence of higher order aberrations, and address inconsistent guidelines regarding the level of aberrations that is acceptable in cryo-EM SPR experiments.
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spelling pubmed-72012892020-05-19 High-resolution cryo-EM reconstructions in the presence of substantial aberrations Bromberg, Raquel Guo, Yirui Borek, Dominika Otwinowski, Zbyszek IUCrJ Research Papers Here, an analysis is performed of how uncorrected antisymmetric aberrations, such as coma and trefoil, affect cryo-EM single-particle reconstruction (SPR) results, and an analytical formula quantifying information loss owing to their presence is inferred that explains why Fourier-shell coefficient-based statistics may report significantly overestimated resolution if these aberrations are not fully corrected. The analysis is validated with reference-based aberration refinement for two cryo-EM SPR data sets acquired with a 200 kV microscope in the presence of coma exceeding 40 µm, and 2.3 and 2.7 Å reconstructions for 144 and 173 kDa particles, respectively, were obtained. The results provide a description of an efficient approach for assessing information loss in cryo-EM SPR data acquired in the presence of higher order aberrations, and address inconsistent guidelines regarding the level of aberrations that is acceptable in cryo-EM SPR experiments. International Union of Crystallography 2020-03-26 /pmc/articles/PMC7201289/ /pubmed/32431828 http://dx.doi.org/10.1107/S2052252520002444 Text en © Raquel Bromberg et al. 2020 http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.http://creativecommons.org/licenses/by/4.0/
spellingShingle Research Papers
Bromberg, Raquel
Guo, Yirui
Borek, Dominika
Otwinowski, Zbyszek
High-resolution cryo-EM reconstructions in the presence of substantial aberrations
title High-resolution cryo-EM reconstructions in the presence of substantial aberrations
title_full High-resolution cryo-EM reconstructions in the presence of substantial aberrations
title_fullStr High-resolution cryo-EM reconstructions in the presence of substantial aberrations
title_full_unstemmed High-resolution cryo-EM reconstructions in the presence of substantial aberrations
title_short High-resolution cryo-EM reconstructions in the presence of substantial aberrations
title_sort high-resolution cryo-em reconstructions in the presence of substantial aberrations
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7201289/
https://www.ncbi.nlm.nih.gov/pubmed/32431828
http://dx.doi.org/10.1107/S2052252520002444
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