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- EMDB-26207: The 3D structure and in situ arrangements (Backslash) of CatSper ... -

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Basic information

Entry
Database: EMDB / ID: EMD-26207
TitleThe 3D structure and in situ arrangements (Backslash) of CatSper channel from the cryo-electron tomography and subtomographic average of mouse efcab9 mutant sperm flagella
Map data
Sample
  • Organelle or cellular component: sperm flagella from efcab9 mutant mouse
Biological speciesMus musculus (house mouse)
Methodsubtomogram averaging / cryo EM / Resolution: 20.7 Å
AuthorsZhao Y / Wang H / Wiesehoefer C / Shah NB / Reetz E / Hwang J / Huang X / Lishko PV / Davies KM / Wennemuth G ...Zhao Y / Wang H / Wiesehoefer C / Shah NB / Reetz E / Hwang J / Huang X / Lishko PV / Davies KM / Wennemuth G / Nicastro D / Chung JJ
Funding support United States, Germany, 6 items
OrganizationGrant numberCountry
National Institutes of Health/Eunice Kennedy Shriver National Institute of Child Health & Human Development (NIH/NICHD)R01HD096745 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R01GM083122 United States
Cancer Prevention and Research Institute of Texas (CPRIT)RR140082 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R01GM111802 United States
Department of Energy (DOE, United States)DE-AC02-O5CH11231 United States
German Research Foundation (DFG)2344/9-3 Germany
CitationJournal: Nat Commun / Year: 2022
Title: 3D structure and in situ arrangements of CatSper channel in the sperm flagellum.
Authors: Yanhe Zhao / Huafeng Wang / Caroline Wiesehoefer / Naman B Shah / Evan Reetz / Jae Yeon Hwang / Xiaofang Huang / Tse-En Wang / Polina V Lishko / Karen M Davies / Gunther Wennemuth / Daniela ...Authors: Yanhe Zhao / Huafeng Wang / Caroline Wiesehoefer / Naman B Shah / Evan Reetz / Jae Yeon Hwang / Xiaofang Huang / Tse-En Wang / Polina V Lishko / Karen M Davies / Gunther Wennemuth / Daniela Nicastro / Jean-Ju Chung /
Abstract: The sperm calcium channel CatSper plays a central role in successful fertilization as a primary Ca gateway. Here, we applied cryo-electron tomography to visualize the higher-order organization of the ...The sperm calcium channel CatSper plays a central role in successful fertilization as a primary Ca gateway. Here, we applied cryo-electron tomography to visualize the higher-order organization of the native CatSper complex in intact mammalian sperm. The repeating CatSper units form long zigzag-rows along mouse and human sperm flagella. Above each tetrameric channel pore, most of the extracellular domains form a canopy that interconnects to a zigzag-shaped roof. Murine CatSper contains an additional wing-structure connected to the tetrameric channel. The intracellular domains link two neighboring channels to a diagonal array, suggesting a dimer formation. Fitting of an atomic model of isolated monomeric CatSper to the in situ map reveals supramolecular interactions and assembly of the CatSper complex. Loss of EFCAB9-CATSPERζ alters the architecture and interactions of the channels, resulting in fragmentation and misalignment of the zigzag-rows and disruption of flagellar movement in Efcab9 sperm. This work offers unique insights into the structural basis for understanding CatSper regulation of sperm motility.
History
DepositionFeb 15, 2022-
Header (metadata) releaseJun 15, 2022-
Map releaseJun 15, 2022-
UpdateJul 6, 2022-
Current statusJul 6, 2022Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_26207.png

Downloads & links

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Map

FileDownload / File: emd_26207.map.gz / Format: CCP4 / Size: 8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
3.15 Å/pix.
x 180 pix.
= 567.18 Å		3.15 Å/pix.
x 130 pix.
= 409.63 Å		3.15 Å/pix.
x 90 pix.
= 283.59 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

generated in cubic-lattice coordinate

Voxel sizeX=Y=Z: 3.151 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 122.0
Minimum - Maximum114.21886 - 130.14508
Average (Standard dev.)120.77538 (±2.6039464)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions13090180
Spacing90130180
CellA: 283.59 Å / B: 409.63 Å / C: 567.18 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : sperm flagella from efcab9 mutant mouse

EntireName: sperm flagella from efcab9 mutant mouse
Components
  • Organelle or cellular component: sperm flagella from efcab9 mutant mouse

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Supramolecule #1: sperm flagella from efcab9 mutant mouse

SupramoleculeName: sperm flagella from efcab9 mutant mouse / type: organelle_or_cellular_component / ID: 1 / Parent: 0
Source (natural)Organism: Mus musculus (house mouse) / Cell: sperm

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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: 7.4
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Specialist opticsPhase plate: VOLTA PHASE PLATE
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 1.75 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 0.5 µm / Nominal defocus min: 0.5 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionApplied symmetry - Point group: C2 (2 fold cyclic) / Algorithm: BACK PROJECTION / Resolution.type: BY AUTHOR / Resolution: 20.7 Å / Resolution method: OTHER
Details: Gold standard but resolution was reported at two cut-offs: 20.7 (0.143), 31.2 (0.5).
Number subtomograms used: 1072
ExtractionNumber tomograms: 19 / Number images used: 536
Final angle assignmentType: OTHER

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