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Title | High-resolution cryo-EM reconstructions in the presence of substantial aberrations. |
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Journal, issue, pages | IUCrJ, Vol. 7, Issue Pt 3, Page 445-452, Year 2020 |
Publish date | May 1, 2020 |
Authors | Raquel Bromberg / Yirui Guo / Dominika Borek / Zbyszek Otwinowski / |
PubMed Abstract | 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 ...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. |
External links | IUCrJ / PubMed:32431828 / PubMed Central |
Methods | EM (single particle) |
Resolution | 2.32 - 2.7 Å |
Structure data | EMDB-21371, PDB-6vrs: EMDB-21373, PDB-6vsa: EMDB-21376, PDB-6vsc: |
Chemicals | ChemComp-MN: ChemComp-HOH: |
Source |
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Keywords | ISOMERASE / glucose isomerase / OXIDOREDUCTASE / heme-binding |