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- EMDB-55045: Capped GJ with additional cytosolic Density -

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Basic information

Entry
Database: EMDB / ID: EMD-55045
TitleCapped GJ with additional cytosolic Density
Map data
Sample
  • Complex: in situ Gap Junction C. elegans
Keywordsin situ Gap Junction ion transfer cellular communication electrical synapse cell-cell contact / MEMBRANE PROTEIN
Biological speciesCaenorhabditis elegans (invertebrata)
Methodsubtomogram averaging / cryo EM / Resolution: 25.6 Å
AuthorsRosenkranz N / Gottschalk A
Funding support Germany, 1 items
OrganizationGrant numberCountry
German Research Foundation (DFG) Germany
CitationJournal: Sci Adv / Year: 2025
Title: In situ structure of a gap junction-stomatin complex.
Authors: Nils Rosenkranz / Alexandra N Birtasu / Konstantin Wieland / Lisa Rehm / Rachita Sharma / Atal Vats / Sina Manger / Aayush Srivastava / Abhishek Bhattacharya / Gerhard Hummer / Achilleas S ...Authors: Nils Rosenkranz / Alexandra N Birtasu / Konstantin Wieland / Lisa Rehm / Rachita Sharma / Atal Vats / Sina Manger / Aayush Srivastava / Abhishek Bhattacharya / Gerhard Hummer / Achilleas S Frangakis / Alexander Gottschalk /
Abstract: Gap junctions (GJs) are intercellular channels that mediate electrical signals and transfer of small molecules. They are crucial for brain, heart, and other organ functions. While molecular ...Gap junctions (GJs) are intercellular channels that mediate electrical signals and transfer of small molecules. They are crucial for brain, heart, and other organ functions. While molecular structures of purified homomeric GJs are available, information of in situ structures is lacking. In vivo, GJs can form heteromers with different functionalities and may associate with other proteins. Here, we analyzed GJs by cryo-electron tomography and subtomogram averaging. We observed hexagonal arrays of GJs at cellular junctions in primary embryonal cells that displayed distinct wide and narrow conformations. Moreover, we found a cap-like, cytosolic protein assembly enclosing the channel pore. We propose that the cap is formed by the stomatin UNC-1, known to interact with UNC-9 innexins. This is corroborated by matching AlphaFold3 models of UNC-1 multimers with our subtomogram average structure; by expressing GFP-tagged UNC-1, leading to cap structures with additional density; and by coarse-grained MD simulations. UNC-1/stomatin rings may affect GJ formation or functions, possibly beyond nematodes.
History
DepositionSep 9, 2025-
Header (metadata) releaseOct 8, 2025-
Map releaseOct 8, 2025-
UpdateNov 26, 2025-
Current statusNov 26, 2025Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_55045.png

Downloads & links

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Map

FileDownload / File: emd_55045.map.gz / Format: CCP4 / Size: 1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
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Others
AxesZ (Sec.)Y (Row.)X (Col.)
5.15 Å/pix.
x 64 pix.
= 329.728 Å		5.15 Å/pix.
x 64 pix.
= 329.728 Å		5.15 Å/pix.
x 64 pix.
= 329.728 Å

Surface

Projections

Slices (1/3)

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Images are generated by Spider.

Voxel sizeX=Y=Z: 5.152 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.95
Minimum - Maximum-7.3020163 - 5.8228536
Average (Standard dev.)0.000000000015414 (±0.9999981)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions646464
Spacing646464
CellA=B=C: 329.728 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_55045_msk_1.map
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Half map: #1

Fileemd_55045_half_map_1.map
Projections & Slices
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Half map: #2

Fileemd_55045_half_map_2.map
Projections & Slices
AxesZYX

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Sample components

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Entire : in situ Gap Junction C. elegans

EntireName: in situ Gap Junction C. elegans
Components
  • Complex: in situ Gap Junction C. elegans

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Supramolecule #1: in situ Gap Junction C. elegans

SupramoleculeName: in situ Gap Junction C. elegans / type: complex / ID: 1 / Parent: 0
Source (natural)Organism: Caenorhabditis elegans (invertebrata)

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation statecell

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Sample preparation

BufferpH: 7
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeTFS KRIOS
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 3.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: OTHER / Nominal defocus max: 3.0 µm / Nominal defocus min: 3.0 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 25.6 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: Super-sampling SART / Number subtomograms used: 336
ExtractionNumber tomograms: 16 / Number images used: 336 / Software - Name: Super-sampling SART
CTF correctionSoftware - Name: CTFFIND / Type: PHASE FLIPPING AND AMPLITUDE CORRECTION
Final angle assignmentType: OTHER / Software - Name: Super-sampling SART

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