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- EMDB-35228: B3 part of axonemal doublet microtubules in mouse sperm with 48-n... -

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Basic information

Entry
Database: EMDB / ID: EMD-35228
TitleB3 part of axonemal doublet microtubules in mouse sperm with 48-nm repeat
Map dataB3 part of axonemal doublet microtubules in mouse sperm with 48-nm repeat
Sample
  • Cell: mouse sperm
Keywordsmicrotubules / axoneme / sperm / filament / PROTEIN TRANSPORT
Biological speciesMus musculus (house mouse)
Methodsubtomogram averaging / cryo EM / Resolution: 7.5 Å
AuthorsZhu Y / Tai LH / Yin GL / Sun F
Funding support China, 1 items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC)31925026 China
CitationJournal: Cell Discov / Year: 2023
Title: In-cell structural insight into the stability of sperm microtubule doublet.
Authors: Linhua Tai / Guoliang Yin / Xiaojun Huang / Fei Sun / Yun Zhu /
Abstract: The propulsion for mammalian sperm swimming is generated by flagella beating. Microtubule doublets (DMTs) along with microtubule inner proteins (MIPs) are essential structural blocks of flagella. ...The propulsion for mammalian sperm swimming is generated by flagella beating. Microtubule doublets (DMTs) along with microtubule inner proteins (MIPs) are essential structural blocks of flagella. However, the intricate molecular architecture of intact sperm DMT remains elusive. Here, by in situ cryo-electron tomography, we solved the in-cell structure of mouse sperm DMT at 4.5-7.5 Å resolutions, and built its model with 36 kinds of MIPs in 48 nm periodicity. We identified multiple copies of Tektin5 that reinforce Tektin bundle, and multiple MIPs with different periodicities that anchor the Tektin bundle to tubulin wall. This architecture contributes to a superior stability of A-tubule than B-tubule of DMT, which was revealed by structural comparison of DMTs from the intact and deformed axonemes. Our work provides an overall molecular picture of intact sperm DMT in 48 nm periodicity that is essential to understand the molecular mechanism of sperm motility as well as the related ciliopathies.
History
DepositionFeb 1, 2023-
Header (metadata) releaseOct 11, 2023-
Map releaseOct 11, 2023-
UpdateDec 6, 2023-
Current statusDec 6, 2023Processing site: PDBj / Status: Released

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Structure visualization

Supplemental images

emd_35228.png

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Map

FileDownload / File: emd_35228.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationB3 part of axonemal doublet microtubules in mouse sperm with 48-nm repeat
Projections & slices

Image control

Size
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AxesZ (Sec.)Y (Row.)X (Col.)
1.76 Å/pix.
x 256 pix.
= 450.56 Å		1.76 Å/pix.
x 256 pix.
= 450.56 Å		1.76 Å/pix.
x 256 pix.
= 450.56 Å

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Images are generated by Spider.

Voxel sizeX=Y=Z: 1.76 Å
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Histogram (log scale)
Contour LevelBy AUTHOR: 0.003
Minimum - Maximum-0.0032472583 - 0.008339567
Average (Standard dev.)0.0002952687 (±0.0015019265)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 450.56 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_35228_msk_1.map
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Half map: half map 1

Fileemd_35228_half_map_1.map
Annotationhalf map 1
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Half map: half map 2

Fileemd_35228_half_map_2.map
Annotationhalf map 2
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Sample components

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Entire : mouse sperm

EntireName: mouse sperm
Components
  • Cell: mouse sperm

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Supramolecule #1: mouse sperm

SupramoleculeName: mouse sperm / type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Mus musculus (house mouse)

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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

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 3.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 5.0 µm / Nominal defocus min: 1.0 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: 7.5 Å / Resolution method: FSC 0.143 CUT-OFF / Number subtomograms used: 17450
ExtractionNumber tomograms: 689 / Number images used: 17450
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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