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- EMDB-7629: Insulin-bound insulin receptor with spot-to-plunge time of 200ms -

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Basic information

Entry
Database: EMDB / ID: EMD-7629
TitleInsulin-bound insulin receptor with spot-to-plunge time of 200ms
Map dataInsulin-bound insulin receptor with spot-to-plunge time of 200ms
Sample
  • Complex: Insulin-bound insulin receptor
Biological speciesHomo sapiens (human)
Methodelectron tomography / cryo EM
AuthorsNoble AJ / Wei H / Dandey VP / Zhang Z / Potter CS / Carragher B
CitationJournal: Nat Methods / Year: 2018
Title: Reducing effects of particle adsorption to the air-water interface in cryo-EM.
Authors: Alex J Noble / Hui Wei / Venkata P Dandey / Zhening Zhang / Yong Zi Tan / Clinton S Potter / Bridget Carragher /
Abstract: Most protein particles prepared in vitreous ice for single-particle cryo-electron microscopy (cryo-EM) are adsorbed to air-water or substrate-water interfaces, which can cause the particles to adopt ...Most protein particles prepared in vitreous ice for single-particle cryo-electron microscopy (cryo-EM) are adsorbed to air-water or substrate-water interfaces, which can cause the particles to adopt preferred orientations. By using a rapid plunge-freezing robot and nanowire grids, we were able to reduce some of the deleterious effects of the air-water interface by decreasing the dwell time of particles in thin liquid films. We demonstrated this by using single-particle cryo-EM and cryo-electron tomography (cryo-ET) to examine hemagglutinin, insulin receptor complex, and apoferritin.
History
DepositionMar 26, 2018-
Header (metadata) releaseApr 4, 2018-
Map releaseApr 4, 2018-
UpdateOct 17, 2018-
Current statusOct 17, 2018Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Solid view (volume rendering)
  • Imaged by UCSF Chimera
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  • Solid view (volume rendering)
  • Imaged by UCSF Chimera
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Supplemental images

emd_7629.png

Downloads & links

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Map

FileDownload / File: emd_7629.map.gz / Format: CCP4 / Size: 2.1 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationInsulin-bound insulin receptor with spot-to-plunge time of 200ms
Voxel sizeX=Y=Z: 2.16423 Å
Density
Minimum - Maximum-1.1096759 - 1.5435286
Average (Standard dev.)0.28197518 (±0.08152928)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin00-213
Dimensions10811134462
Spacing11341081462
CellA: 2454.237 Å / B: 2339.5327 Å / C: 999.8743 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z2.16423015873022.16423034227572.1642294372294
M x/y/z11341081462
origin x/y/z0.0000.0000.000
length x/y/z2454.2372339.533999.874
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ450450450
MAP C/R/S123
start NC/NR/NS00-213
NC/NR/NS11341081462
D min/max/mean-1.1101.5440.282

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Supplemental data

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Sample components

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Entire : Insulin-bound insulin receptor

EntireName: Insulin-bound insulin receptor
Components
  • Complex: Insulin-bound insulin receptor

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Supramolecule #1: Insulin-bound insulin receptor

SupramoleculeName: Insulin-bound insulin receptor / type: complex / ID: 1 / Parent: 0
Details: Insulin Receptor Ectodomain in complex with two insulin molecules. Complete sample details may be found in EMD-7462.
Source (natural)Organism: Homo sapiens (human)

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation stateparticle

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Sample preparation

BufferpH: 7.5
GridModel: Homemade
VitrificationCryogen name: ETHANE / Instrument: OTHER
Details: The value given for _emd_vitrification.instrument is SPOTITON. This is not in a list of allowed values set(['LEICA EM CPC', 'GATAN CRYOPLUNGE 3', 'LEICA PLUNGER', 'FEI VITROBOT MARK II', ...Details: The value given for _emd_vitrification.instrument is SPOTITON. This is not in a list of allowed values set(['LEICA EM CPC', 'GATAN CRYOPLUNGE 3', 'LEICA PLUNGER', 'FEI VITROBOT MARK II', 'HOMEMADE PLUNGER', 'REICHERT-JUNG PLUNGER', 'FEI VITROBOT MARK I', 'LEICA KF80', 'FEI VITROBOT MARK III', 'LEICA EM GP', 'OTHER', 'FEI VITROBOT MARK IV']) so OTHER is written into the XML file.
SectioningOther: NO SECTIONING

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy
Specialist opticsEnergy filter - Name: GIF Bioquantum
Image recordingFilm or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: COUNTING / Average electron dose: 3.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionAlgorithm: SIMULTANEOUS ITERATIVE (SIRT) / Software - Name: TOMO3D / Software - details: SIRT, 30 iterations / Number images used: 30

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