[English] 日本語
Yorodumi
- EMDB-37276: Local refinement cryo-EM map of human 26S RP (Ea state) bound to ... -

+
Open data


ID or keywords:

Loading...

-
Basic information

Entry
Database: EMDB / ID: EMD-37276
TitleLocal refinement cryo-EM map of human 26S RP (Ea state) bound to K11/K48-branched ubiquitin (Ub) chain composed of three Ub, focused on Rpn3/Rpn7 region.
Map dataLocal refinement cryo-EM map of human 26S RP (Ea state) bound to K11/K48-branched ubiquitin (Ub) chain composed of three Ub, focused on Rpn3/Rpn7 region.
Sample
  • Complex: Human 26S proteasome
    • Complex: 19S regulatory particle (RP) of human 26S proteasome
      • Complex: AAA+-ATPase of 19S regulatory particle (RP) of human 26S proteasome
    • Complex: alpha subunits belonging to the 20S core particle (CP) of human 26S proteasome
    • Complex: K11/K48-branched ubiquitin (Ub) chain composed of three Ub
Keywordsprotein degradation / macromolecular complex / ubiquitin-proteasome system / CYTOSOLIC PROTEIN
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.0 Å
AuthorsHsu STD / Draczkowski P / Wang YS
Funding support Taiwan, 4 items
OrganizationGrant numberCountry
Academia Sinica (Taiwan)AS-CDA-109- L08 Taiwan
Ministry of Science and Technology (MoST, Taiwan)MOST 109-3114-Y-001-001 Taiwan
Ministry of Science and Technology (MoST, Taiwan)MOST 110-2113-M-001- 050-MY3 Taiwan
Ministry of Science and Technology (MoST, Taiwan)MOST 110-2311-B-001-013-MY3 Taiwan
CitationJournal: To Be Published
Title: Cryo-EM structure of human 26S proteasomal RP subcomplex bound with branched Ub chain.
Authors: Draczkowski P / Hsu STD / Wang YS
History
DepositionAug 26, 2023-
Header (metadata) releaseAug 28, 2024-
Map releaseAug 28, 2024-
UpdateAug 28, 2024-
Current statusAug 28, 2024Processing site: PDBj / Status: Released

-
Structure visualization

Supplemental images

emd_37276.png

Downloads & links

-
Map

FileDownload / File: emd_37276.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationLocal refinement cryo-EM map of human 26S RP (Ea state) bound to K11/K48-branched ubiquitin (Ub) chain composed of three Ub, focused on Rpn3/Rpn7 region.
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.4 Å/pix.
x 400 pix.
= 560. Å		1.4 Å/pix.
x 400 pix.
= 560. Å		1.4 Å/pix.
x 400 pix.
= 560. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.4 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.7
Minimum - Maximum-0.40324315 - 1.5792775
Average (Standard dev.)-0.002364333 (±0.042403474)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 560.0 Å
α=β=γ: 90.0 °

-
Supplemental data

-
Half map: Local refinement cryo-EM half map of human 26S...

Fileemd_37276_half_map_1.map
AnnotationLocal refinement cryo-EM half map of human 26S RP (Ea state) bound to K11/K48-branched ubiquitin (Ub) chain composed of three Ub, focused on Rpn3/Rpn7 region.
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

-
Half map: Local refinement cryo-EM half map of human 26S...

Fileemd_37276_half_map_2.map
AnnotationLocal refinement cryo-EM half map of human 26S RP (Ea state) bound to K11/K48-branched ubiquitin (Ub) chain composed of three Ub, focused on Rpn3/Rpn7 region.
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

-
Sample components

-
Entire : Human 26S proteasome

EntireName: Human 26S proteasome
Components
  • Complex: Human 26S proteasome
    • Complex: 19S regulatory particle (RP) of human 26S proteasome
      • Complex: AAA+-ATPase of 19S regulatory particle (RP) of human 26S proteasome
    • Complex: alpha subunits belonging to the 20S core particle (CP) of human 26S proteasome
    • Complex: K11/K48-branched ubiquitin (Ub) chain composed of three Ub

-
Supramolecule #1: Human 26S proteasome

SupramoleculeName: Human 26S proteasome / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#26
Details: Images of double-capped (RP-CP-RP) 26S proteasome particles were split in half. After combining with subset of single-capped (RP-CP) complexes the signal in particle images was partially ...Details: Images of double-capped (RP-CP-RP) 26S proteasome particles were split in half. After combining with subset of single-capped (RP-CP) complexes the signal in particle images was partially subtracted leaving only the signal of 19S RP together with alpha subunits of the 20S CP proteasomal subcomplex.
Source (natural)Organism: Homo sapiens (human)

-
Supramolecule #2: 19S regulatory particle (RP) of human 26S proteasome

SupramoleculeName: 19S regulatory particle (RP) of human 26S proteasome / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1-#4, #12-#25
Source (natural)Organism: Homo sapiens (human)

-
Supramolecule #3: AAA+-ATPase of 19S regulatory particle (RP) of human 26S proteasome

SupramoleculeName: AAA+-ATPase of 19S regulatory particle (RP) of human 26S proteasome
type: complex / ID: 3 / Parent: 2 / Macromolecule list: #1-#4, #24-#25
Source (natural)Organism: Homo sapiens (human)

-
Supramolecule #4: alpha subunits belonging to the 20S core particle (CP) of human 2...

SupramoleculeName: alpha subunits belonging to the 20S core particle (CP) of human 26S proteasome
type: complex / ID: 4 / Parent: 1 / Macromolecule list: #5-#11
Source (natural)Organism: Homo sapiens (human)

-
Supramolecule #5: K11/K48-branched ubiquitin (Ub) chain composed of three Ub

SupramoleculeName: K11/K48-branched ubiquitin (Ub) chain composed of three Ub
type: complex / ID: 5 / Parent: 1 / Macromolecule list: #26
Source (natural)Organism: Homo sapiens (human)

-
Experimental details

-
Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

-
Sample preparation

Concentration1 mg/mL
BufferpH: 7.6
Component:
ConcentrationFormulaName
20.0 mMC8H18N2O4SHEPES
40.0 mMNaClsodium chloride
5.0 mMMgCl2magnesium chloride
2.0 mMC2H6OS2-Mercaptoethanol
0.5 mMC12H8N21,10-Phenanthroline
2.0 mMC10H16N5O13P3adenosine triphosphate
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR / Details: 20 mA
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV / Details: incubation time= 3 s blotting time= 2.5 s.
DetailsThe complex was additionally supplemented with an excess of preformed and SEC-purified RPN13:UCHL5 complex.

-
Electron microscopy

MicroscopeFEI TITAN KRIOS
Specialist opticsEnergy filter - Slit width: 20 eV
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 3 / Number real images: 15242 / Average exposure time: 1.8 sec. / Average electron dose: 49.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.8 µm / Nominal defocus min: 1.2 µm / Nominal magnification: 70000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

+
Image processing

Particle selectionNumber selected: 2695911
Startup modelType of model: INSILICO MODEL
In silico model: Generated by cryoSPARC using Ab-initio Reconstruction job type.
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 4.0 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 3.1) / Software - details: Non-uniform Refinment / Number images used: 54794
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 3.1) / Software - details: Non-uniform Refinment
Details: Performed in cryoSPARC using Local focused refinement.
Final 3D classificationNumber classes: 6 / Software - Name: cryoSPARC (ver. 3.1)
Details: 3D variability analysis (3DVA) followed by Heterogenous refinement using 3DVA cluster volumes as starting references.

+
About Yorodumi

-
News

-
Feb 9, 2022. New format data for meta-information of EMDB entries

New format data for meta-information of EMDB entries

  • Version 3 of the EMDB header file is now the official format.
  • The previous official version 1.9 will be removed from the archive.

Related info.:EMDB header

External links:wwPDB to switch to version 3 of the EMDB data model

-
Aug 12, 2020. Covid-19 info

Covid-19 info

URL: https://pdbj.org/emnavi/covid19.php

New page: Covid-19 featured information page in EM Navigator.

Related info.:Covid-19 info / Mar 5, 2020. Novel coronavirus structure data

+
Mar 5, 2020. Novel coronavirus structure data

Novel coronavirus structure data

Related info.:Yorodumi Speices / Aug 12, 2020. Covid-19 info

External links:COVID-19 featured content - PDBj / Molecule of the Month (242)：Coronavirus Proteases

+
Jan 31, 2019. EMDB accession codes are about to change! (news from PDBe EMDB page)

EMDB accession codes are about to change! (news from PDBe EMDB page)

  • The allocation of 4 digits for EMDB accession codes will soon come to an end. Whilst these codes will remain in use, new EMDB accession codes will include an additional digit and will expand incrementally as the available range of codes is exhausted. The current 4-digit format prefixed with “EMD-” (i.e. EMD-XXXX) will advance to a 5-digit format (i.e. EMD-XXXXX), and so on. It is currently estimated that the 4-digit codes will be depleted around Spring 2019, at which point the 5-digit format will come into force.
  • The EM Navigator/Yorodumi systems omit the EMD- prefix.

Related info.:Q: What is EMD? / ID/Accession-code notation in Yorodumi/EM Navigator

External links:EMDB Accession Codes are Changing Soon! / Contact to PDBj

+
Jul 12, 2017. Major update of PDB

Major update of PDB

  • wwPDB released updated PDB data conforming to the new PDBx/mmCIF dictionary.
  • This is a major update changing the version number from 4 to 5, and with Remediation, in which all the entries are updated.
  • In this update, many items about electron microscopy experimental information are reorganized (e.g. em_software).
  • Now, EM Navigator and Yorodumi are based on the updated data.

External links:wwPDB Remediation / Enriched Model Files Conforming to OneDep Data Standards Now Available in the PDB FTP Archive

-
Yorodumi

Thousand views of thousand structures

  • Yorodumi is a browser for structure data from EMDB, PDB, SASBDB, etc.
  • This page is also the successor to EM Navigator detail page, and also detail information page/front-end page for Omokage search.
  • The word "yorodu" (or yorozu) is an old Japanese word meaning "ten thousand". "mi" (miru) is to see.

Related info.:EMDB / PDB / SASBDB / Comparison of 3 databanks / Yorodumi Search / Aug 31, 2016. New EM Navigator & Yorodumi / Yorodumi Papers / Jmol/JSmol / Function and homology information / Changes in new EM Navigator and Yorodumi

Read more