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- EMDB-9233: T20S proteasome -

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Basic information

Entry
Database: EMDB / ID: EMD-9233
TitleT20S proteasome
Map dataT20S sharpened masked map
Sample
  • Complex: 20S proteasomeProteasome
Biological speciesThermoplasma acidophilum (acidophilic)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.1 Å
AuthorsEng ET / Kopylov M / Jordan KJ / Rice WJ / Carragher BO / Potter CS
CitationJournal: J Struct Biol / Year: 2019
Title: Reducing cryoEM file storage using lossy image formats.
Authors: Edward T Eng / Mykhailo Kopylov / Carl J Negro / Sarkis Dallaykan / William J Rice / Kelsey D Jordan / Kotaro Kelley / Bridget Carragher / Clinton S Potter /
Abstract: Recent advances in instrumentation and software for cryoEM have increased the applicability and utility of this method. High levels of automation and faster data acquisition rates require hard ...Recent advances in instrumentation and software for cryoEM have increased the applicability and utility of this method. High levels of automation and faster data acquisition rates require hard decisions to be made regarding data retention. Here we investigate the efficacy of data compression applied to aligned summed movie files. Surprisingly, these images can be compressed using a standard lossy method that reduces file storage by 90-95% and yet can still be processed to provide sub-2 Å reconstructed maps. We do not advocate this as an archival method, but it may provide a useful means for retaining images as an historical record, especially at large facilities.
History
DepositionOct 18, 2018-
Header (metadata) releaseOct 31, 2018-
Map releaseOct 31, 2018-
UpdateOct 31, 2018-
Current statusOct 31, 2018Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.02
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by cylindrical radius
  • Surface level: 0.02
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_9233.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationT20S sharpened masked map
Voxel sizeX=Y=Z: 0.6616 Å
Density
Contour LevelBy AUTHOR: 0.02 / Movie #1: 0.02
Minimum - Maximum-0.08436498 - 0.14498375
Average (Standard dev.)0.00018564082 (±0.004726364)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 264.63998 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z0.66160.66160.6616
M x/y/z400400400
origin x/y/z0.0000.0000.000
length x/y/z264.640264.640264.640
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS400400400
D min/max/mean-0.0840.1450.000

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Supplemental data

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Mask #1

Fileemd_9233_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Additional map: T20S unsharpened map

Fileemd_9233_additional.map
AnnotationT20S unsharpened map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: T20S even half map

Fileemd_9233_half_map_1.map
AnnotationT20S even half map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: T20S odd half map

Fileemd_9233_half_map_2.map
AnnotationT20S odd half map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : 20S proteasome

EntireName: 20S proteasomeProteasome
Components
  • Complex: 20S proteasomeProteasome

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Supramolecule #1: 20S proteasome

SupramoleculeName: 20S proteasome / type: complex / ID: 1 / Parent: 0
Source (natural)Organism: Thermoplasma acidophilum (acidophilic)
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria) / Recombinant plasmid: pREAR-A
Molecular weightTheoretical: 700 KDa

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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

Concentration0.21 mg/mL
BufferpH: 7.4
Component:
ConcentrationFormulaName
150.0 mMNaClSodium chloridesodium chloride
20.0 mMC4H11NO3Tris

Details: 20 mM Tris, 150 mM NaCl
GridModel: C-flat-1.2/1.3 4C / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY ARRAY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: OTHER
VitrificationCryogen name: ETHANE / Chamber humidity: 75 % / Chamber temperature: 298 K / Instrument: GATAN CRYOPLUNGE 3
Details: Blotted for 2.5 seconds after a 30 second wait time..
Details1.2/1.3 C-Flat grid, plasma cleaned

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 70.0 µm / Calibrated defocus max: 3.02 µm / Calibrated defocus min: 0.2 µm / Calibrated magnification: 75574 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 37000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
TemperatureMin: 70.0 K / Max: 70.0 K
DetailsLeginon
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3710 pixel / Digitization - Dimensions - Height: 3838 pixel / Digitization - Sampling interval: 5.0 µm / Digitization - Frames/image: 1-50 / Number grids imaged: 1 / Number real images: 1173 / Average exposure time: 6.259 sec. / Average electron dose: 62.59 e/Å2
Details: Images were collected in movie mode at 5 frames per second.
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 172896
Details: Template picking by FindEM. Templates were created from Appion DoG picks from the first 50 images, which were subjected to 2D classification in RELION.
CTF correctionSoftware - Name: CTFFIND (ver. 4)
Details: CTF amplitude correction was performed following 3D reconstruction.
Startup modelType of model: INSILICO MODEL / In silico model: Ab initio model created by cryoSPARC
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3)
Final 3D classificationNumber classes: 3 / Software - Name: RELION (ver. 3)
Details: 3D classification used K=3 and 1 out of the 3 classes were chosen for the final refinement.
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3)
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: D7 (2x7 fold dihedral) / Algorithm: BACK PROJECTION / Resolution.type: BY AUTHOR / Resolution: 2.1 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3) / Number images used: 127570
DetailsImages were automatically run through MotionCor2 by Appion on-the-fly with the aligned sum and aligned dose-weighted sum saved.
FSC plot (resolution estimation)

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