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- EMDB-10840: Structure of RyR1 in apo/closed state in native membrane -

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Basic information

Entry
Database: EMDB / ID: EMD-10840
TitleStructure of RyR1 in apo/closed state in native membrane
Map data
SampleSarcoplasmic reticulum
Function / homology
Function and homology information


integral component of organelle membrane / ATP-gated ion channel activity / ryanodine-sensitive calcium-release channel activity / terminal cisterna / ryanodine receptor complex / release of sequestered calcium ion into cytosol by sarcoplasmic reticulum / ossification involved in bone maturation / cellular response to caffeine / calcium-release channel activity / skin development ...integral component of organelle membrane / ATP-gated ion channel activity / ryanodine-sensitive calcium-release channel activity / terminal cisterna / ryanodine receptor complex / release of sequestered calcium ion into cytosol by sarcoplasmic reticulum / ossification involved in bone maturation / cellular response to caffeine / calcium-release channel activity / skin development / outflow tract morphogenesis / voltage-gated calcium channel activity / skeletal muscle fiber development / release of sequestered calcium ion into cytosol / toxic substance binding / cellular response to calcium ion / calcium ion transmembrane transport / calcium channel activity / sarcoplasmic reticulum membrane / sarcoplasmic reticulum / muscle contraction / disordered domain specific binding / drug binding / ion channel binding / protein homotetramerization / calmodulin binding / calcium ion binding / membrane / integral component of membrane / ATP binding / identical protein binding
MIR motif / Ryanodine receptor, SPRY domain 2 / RIH domain / B30.2/SPRY domain / Ryanodine receptor 1 / Ryanodine receptor Ryr / Ryanodine receptor, SPRY domain 1 / SPRY domain / Ion transport domain / Ryanodine receptor, SPRY domain 3 ...MIR motif / Ryanodine receptor, SPRY domain 2 / RIH domain / B30.2/SPRY domain / Ryanodine receptor 1 / Ryanodine receptor Ryr / Ryanodine receptor, SPRY domain 1 / SPRY domain / Ion transport domain / Ryanodine receptor, SPRY domain 3 / Ryanodine Receptor TM 4-6 / EF-hand domain pair / Concanavalin A-like lectin/glucanase domain superfamily / Ryanodine receptor / RyR/IP3 receptor binding core, RIH domain superfamily / Mir domain superfamily / B30.2/SPRY domain superfamily / RyR/IP3R Homology associated domain / Inositol 1,4,5-trisphosphate/ryanodine receptor
Ryanodine receptor 1
Biological speciesOryctolagus cuniculus (rabbit)
Methodsubtomogram averaging / cryo EM / Resolution: 9.1 Å
AuthorsChen W / Sanchez R / Zhang Y / Dietrich L / Kudryashev M
Funding support Germany, 1 items
OrganizationGrant numberCountry
Alexander von Humboldt FoundationSofja Kovalevskaja Award Germany
CitationJournal: Nat Commun / Year: 2020
Title: Subnanometer-resolution structure determination in situ by hybrid subtomogram averaging - single particle cryo-EM.
Authors: Ricardo M Sanchez / Yingyi Zhang / Wenbo Chen / Lea Dietrich / Mikhail Kudryashev /
Abstract: Cryo-electron tomography combined with subtomogram averaging (StA) has yielded high-resolution structures of macromolecules in their native context. However, high-resolution StA is not commonplace ...Cryo-electron tomography combined with subtomogram averaging (StA) has yielded high-resolution structures of macromolecules in their native context. However, high-resolution StA is not commonplace due to beam-induced sample drift, images with poor signal-to-noise ratios (SNR), challenges in CTF correction, and limited particle number. Here we address these issues by collecting tilt series with a higher electron dose at the zero-degree tilt. Particles of interest are then located within reconstructed tomograms, processed by conventional StA, and then re-extracted from the high-dose images in 2D. Single particle analysis tools are then applied to refine the 2D particle alignment and generate a reconstruction. Use of our hybrid StA (hStA) workflow improved the resolution for tobacco mosaic virus from 7.2 to 4.4 Å and for the ion channel RyR1 in crowded native membranes from 12.9 to 9.1 Å. These resolution gains make hStA a promising approach for other StA projects aimed at achieving subnanometer resolution.
History
DepositionApr 7, 2020-
Header (metadata) releaseAug 5, 2020-
Map releaseAug 5, 2020-
UpdateAug 5, 2020-
Current statusAug 5, 2020Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.0227
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by cylindrical radius
  • Surface level: 0.0227
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_10840.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.7 Å/pix.
x 320 pix.
= 544. Å
1.7 Å/pix.
x 320 pix.
= 544. Å
1.7 Å/pix.
x 320 pix.
= 544. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.7 Å
Density
Contour LevelBy AUTHOR: 0.0227 / Movie #1: 0.0227
Minimum - Maximum-0.05186503 - 0.0947685
Average (Standard dev.)0.0004150545 (±0.00484771)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions320320320
Spacing320320320
CellA=B=C: 544.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.71.71.7
M x/y/z320320320
origin x/y/z0.0000.0000.000
length x/y/z544.000544.000544.000
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS320320320
D min/max/mean-0.0520.0950.000

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Supplemental data

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Segmentation: #1

Fileemd_10840_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: half map 1 of structure of RyR1 in...

Fileemd_10840_half_map_1.map
Annotationhalf map 1 of structure of RyR1 in native membrane in apo/closed state
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: half map 2 of structure of RyR1 in...

Fileemd_10840_half_map_2.map
Annotationhalf map 2 of structure of RyR1 in native membrane in apo/closed state
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire Sarcoplasmic reticulum

EntireName: Sarcoplasmic reticulum / Number of components: 1

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Component #1: cellular-component, Sarcoplasmic reticulum

Cellular-componentName: Sarcoplasmic reticulum / Recombinant expression: No
SourceSpecies: Oryctolagus cuniculus (rabbit)

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Experimental details

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Sample preparation

SpecimenSpecimen state: Tissue / Method: cryo EM
Sample solutionpH: 7.1
VitrificationCryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
ImagingMicroscope: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 2 e/Å2 / Illumination mode: FLOOD BEAM
LensImaging mode: BRIGHT FIELD
Specimen HolderModel: OTHER
CameraDetector: GATAN K2 SUMMIT (4k x 4k)

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Image processing

ProcessingMethod: subtomogram averaging / Applied symmetry: C4 (4 fold cyclic) / Number of subtomograms: 2563
3D reconstructionResolution: 9.1 Å / Resolution method: FSC 0.143 CUT-OFF

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