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- EMDB-71283: SPOP 14mer Double Donut -

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Open data


ID or keywords:

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Basic information

Entry
Database: EMDB / ID: EMD-71283
TitleSPOP 14mer Double Donut
Map data
Sample
  • Complex: SPOP 14mer double donut
Keywordsubiquitination / protein degradation / protein oligomer / substrate adapter / PROTEIN BINDING
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 11.3 Å
AuthorsCuneo MJ
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS) United States
CitationJournal: bioRxiv / Year: 2025
Title: The equilibrium between two quaternary assembly states determines the activity of SPOP and its cancer mutants.
Authors: Matthew J Cuneo / Ömer Güllülü / Mohamed-Raafet Ammar / Xinrui Gui / Kelly Churion / Martin Turk / Brian G O'Flynn / Nafiseh Sabri / Tanja Mittag /
Abstract: Proteostasis is critical for preventing oncogenesis. Both activating and inactivating mutations in the ubiquitin ligase subunit SPOP result in oncogenesis in different tissues. SPOP assembles into ...Proteostasis is critical for preventing oncogenesis. Both activating and inactivating mutations in the ubiquitin ligase subunit SPOP result in oncogenesis in different tissues. SPOP assembles into filaments that are multivalent for substrates, and substrates have multiple weak motifs for SPOP that are not activated via post-translational modifications. It is thus unclear how regulation is achieved. Here, we show that SPOP filaments circularize into rings that dimerize into up to 2.5 MDa-large, auto-inhibited double donuts. The equilibrium between double donuts and linear filaments determines SPOP activity. Activating and deactivating cancer mutations shift the equilibrium towards the filament or the double donut, respectively, and this influences substrate turnover and subcellular localization. This regulatory mechanism requires long filaments that can circularize into rings, likely explaining the presence of multiple weak SPOP-binding motifs in substrates. Activating and deactivating mutations combine to give rise to intermediate activities, suggesting new levers for cancer therapies.
History
DepositionJun 17, 2025-
Header (metadata) releaseJul 30, 2025-
Map releaseJul 30, 2025-
UpdateJul 30, 2025-
Current statusJul 30, 2025Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_71283.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
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Others
AxesZ (Sec.)Y (Row.)X (Col.)
2.6 Å/pix.
x 320 pix.
= 832. Å
2.6 Å/pix.
x 320 pix.
= 832. Å
2.6 Å/pix.
x 320 pix.
= 832. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 2.6 Å
Density
Contour LevelBy AUTHOR: 0.35
Minimum - Maximum-0.42457795 - 3.3467581
Average (Standard dev.)-0.00041031025 (±0.11995082)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions320320320
Spacing320320320
CellA=B=C: 832.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_71283_msk_1.map
Projections & Slices
AxesZYX

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Half map: #1

Fileemd_71283_half_map_1.map
Projections & Slices
AxesZYX

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Half map: #2

Fileemd_71283_half_map_2.map
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Sample components

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Entire : SPOP 14mer double donut

EntireName: SPOP 14mer double donut
Components
  • Complex: SPOP 14mer double donut

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Supramolecule #1: SPOP 14mer double donut

SupramoleculeName: SPOP 14mer double donut / type: complex / ID: 1 / Parent: 0
Source (natural)Organism: Homo sapiens (human)

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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

BufferpH: 7.5
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TALOS ARCTICA
Image recordingFilm or detector model: FEI FALCON III (4k x 4k) / Average electron dose: 1.0 e/Å2
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.8000000000000003 µm / Nominal defocus min: 1.0 µm
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

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Image processing

CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: NONE
Final reconstructionResolution.type: BY AUTHOR / Resolution: 11.3 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC / Number images used: 18500
Initial angle assignmentType: NOT APPLICABLE
Final angle assignmentType: NOT APPLICABLE
FSC plot (resolution estimation)

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