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- EMDB-35828: Cryo-EPty SPA at CSA of 1.03 mrad -

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Basic information

Entry
Database: EMDB / ID: EMD-35828
TitleCryo-EPty SPA at CSA of 1.03 mrad
Map dataCryo-EPty SPA 3D map reconstructed from 232 particles at convergence angle of 1.03 mrad
SampleRotavirus A != Rotavirus

Rotavirus A

  • Virus: Rotavirus
KeywordsComplex / VIRUS
Biological speciesRotavirus
Methodsingle particle reconstruction / cryo EM / Resolution: 37.2 Å
AuthorsPei X / Wang P
Funding support China, United Kingdom, European Union, 4 items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC)11874199 China
Medical Research Council (MRC, United Kingdom)MR/N00065X/1 United Kingdom
Wellcome Trust206422/Z/17/Z United Kingdom
European Research Council (ERC)101021133European Union
CitationJournal: Nat Commun / Year: 2023
Title: Cryogenic electron ptychographic single particle analysis with wide bandwidth information transfer.
Authors: Xudong Pei / Liqi Zhou / Chen Huang / Mark Boyce / Judy S Kim / Emanuela Liberti / Yiming Hu / Takeo Sasaki / Peter D Nellist / Peijun Zhang / David I Stuart / Angus I Kirkland / Peng Wang /
Abstract: Advances in cryogenic transmission electron microscopy have revolutionised the determination of many macromolecular structures at atomic or near-atomic resolution. This method is based on ...Advances in cryogenic transmission electron microscopy have revolutionised the determination of many macromolecular structures at atomic or near-atomic resolution. This method is based on conventional defocused phase contrast imaging. However, it has limitations of weaker contrast for small biological molecules embedded in vitreous ice, in comparison with cryo-ptychography, which shows increased contrast. Here we report a single-particle analysis based on the use of ptychographic reconstruction data, demonstrating that three dimensional reconstructions with a wide information transfer bandwidth can be recovered by Fourier domain synthesis. Our work suggests future applications in otherwise challenging single particle analyses, including small macromolecules and heterogeneous or flexible particles. In addition structure determination in situ within cells without the requirement for protein purification and expression may be possible.
History
DepositionApr 5, 2023-
Header (metadata) releaseJun 14, 2023-
Map releaseJun 14, 2023-
UpdateJun 14, 2023-
Current statusJun 14, 2023Processing site: PDBj / Status: Released

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Structure visualization

Supplemental images

emd_35828.png

Downloads & links

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Map

FileDownload / File: emd_35828.map.gz / Format: CCP4 / Size: 58.2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationCryo-EPty SPA 3D map reconstructed from 232 particles at convergence angle of 1.03 mrad
Voxel sizeX=Y=Z: 4.65 Å
Density
Contour LevelBy AUTHOR: 0.657
Minimum - Maximum0.0 - 1.0
Average (Standard dev.)0.4007126 (±0.12844917)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions248248248
Spacing248248248
CellA=B=C: 1153.2001 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #1

Fileemd_35828_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #2

Fileemd_35828_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Rotavirus A

EntireName: Rotavirus A
Components
  • Virus: Rotavirus

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Supramolecule #1: Rotavirus

SupramoleculeName: Rotavirus / type: virus / ID: 1 / Parent: 0 / NCBI-ID: 10912 / Sci species name: Rotavirus / Virus type: VIRUS-LIKE PARTICLE / Virus isolate: STRAIN / Virus enveloped: Yes / Virus empty: No

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration8 mg/mL
BufferpH: 8
GridModel: Quantifoil
VitrificationCryogen name: ETHANE / Chamber humidity: 80 % / Chamber temperature: 298 K / Instrument: GATAN CRYOPLUNGE 3

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Electron microscopy

MicroscopeJEOL CRYO ARM 300
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: OTHER / Nominal defocus max: 14.0 µm / Nominal defocus min: 13.5 µm
Image recordingFilm or detector model: OTHER / Average electron dose: 22.6 e/Å2

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Image processing

Particle selectionNumber selected: 257
Startup modelType of model: INSILICO MODEL
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
Final 3D classificationNumber classes: 3 / Avg.num./class: 86 / Software - Name: RELION (ver. 3.1)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
Final reconstructionNumber classes used: 2 / Applied symmetry - Point group: I (icosahedral) / Resolution.type: BY AUTHOR / Resolution: 37.2 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.1) / Number images used: 232
FSC plot (resolution estimation)

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