Low-cooling-rate freezing in biomolecular cryo-electron microscopy for recovery of initial frames.
beam-induced motion
cooling rate
cryo-EM sample preparation
high temperature freezing
Journal
QRB discovery
ISSN: 2633-2892
Titre abrégé: QRB Discov
Pays: England
ID NLM: 101772102
Informations de publication
Date de publication:
2021
2021
Historique:
received:
18
06
2021
revised:
20
08
2021
accepted:
01
09
2021
medline:
6
9
2021
pubmed:
6
9
2021
entrez:
2
8
2023
Statut:
epublish
Résumé
When biological samples are first exposed to electrons in cryo-electron microcopy (cryo-EM), proteins exhibit a rapid 'burst' phase of beam-induced motion that cannot be corrected with software. This lowers the quality of the initial frames, which are the least damaged by the electrons. Hence, they are commonly excluded or down-weighted during data processing, reducing the undamaged signal and the resolution in the reconstruction. By decreasing the cooling rate during sample preparation, either with a cooling-rate gradient or by increasing the freezing temperature, we show that the quality of the initial frames for various protein and virus samples can be recovered. Incorporation of the initial frames in the reconstruction increases the resolution by an amount equivalent to using ~60% more data. Moreover, these frames preserve the high-quality cryo-EM densities of radiation-sensitive residues, which is often damaged or very weak in canonical three-dimensional reconstruction. The improved freezing conditions can be easily achieved using existing devices and enhance the overall quality of cryo-EM structures.
Identifiants
pubmed: 37529673
doi: 10.1017/qrd.2021.8
pii: S2633289221000089
pmc: PMC10392635
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e11Informations de copyright
© The Author(s) 2021.
Déclaration de conflit d'intérêts
The authors declare no conflict of interest.
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