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Yorodumi- EMDB-41431: 48-nm repeating structure of doublets from mouse sperm flagella -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-41431 | ||||||||||||
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Title | 48-nm repeating structure of doublets from mouse sperm flagella | ||||||||||||
Map data | Final map of 48 nm-repeating unit of mouse sperm doublets | ||||||||||||
Sample |
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Keywords | Mammalian sperm / axoneme / microtubule-based structure / microtubule inner protein / non-motor proteins / cellular motility / fertility / structural protein | ||||||||||||
Function / homology | Function and homology information ERKs are inactivated / left/right pattern formation / epithelial cilium movement involved in determination of left/right asymmetry / sperm flagellum assembly / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Sealing of the nuclear envelope (NE) by ESCRT-III / establishment of left/right asymmetry / 9+0 motile cilium / outer acrosomal membrane ...ERKs are inactivated / left/right pattern formation / epithelial cilium movement involved in determination of left/right asymmetry / sperm flagellum assembly / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Sealing of the nuclear envelope (NE) by ESCRT-III / establishment of left/right asymmetry / 9+0 motile cilium / outer acrosomal membrane / regulation of brood size / axonemal microtubule doublet inner sheath / protein localization to motile cilium / manchette assembly / axonemal B tubule inner sheath / axonemal A tubule inner sheath / inner dynein arm assembly / Intraflagellar transport / regulation of calcineurin-NFAT signaling cascade / Carboxyterminal post-translational modifications of tubulin / MAP kinase phosphatase activity / sperm axoneme assembly / protein polyglutamylation / positive regulation of feeding behavior / sperm principal piece / COPI-independent Golgi-to-ER retrograde traffic / cerebrospinal fluid circulation / microtubule sliding / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / epithelial cilium movement involved in extracellular fluid movement / regulation of store-operated calcium entry / cilium movement involved in cell motility / regulation of cilium beat frequency involved in ciliary motility / protein tyrosine/serine/threonine phosphatase activity / COPI-mediated anterograde transport / Aggrephagy / 9+2 motile cilium / Kinesins / Transferases; Transferring phosphorus-containing groups / intraciliary transport / acrosomal membrane / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / PKR-mediated signaling / Resolution of Sister Chromatid Cohesion / ciliary transition zone / RHO GTPases activate IQGAPs / cilium movement / The role of GTSE1 in G2/M progression after G2 checkpoint / calcium ion sensor activity / axoneme assembly / Recycling pathway of L1 / cilium organization / left/right axis specification / organelle transport along microtubule / axonemal microtubule / COPI-dependent Golgi-to-ER retrograde traffic / RHO GTPases Activate Formins / Separation of Sister Chromatids / gamma-tubulin ring complex / Hedgehog 'off' state / Loss of Nlp from mitotic centrosomes / Recruitment of mitotic centrosome proteins and complexes / Loss of proteins required for interphase microtubule organization from the centrosome / Recruitment of NuMA to mitotic centrosomes / Anchoring of the basal body to the plasma membrane / AURKA Activation by TPX2 / forebrain morphogenesis / negative regulation of T cell activation / glial cell differentiation / Regulation of PLK1 Activity at G2/M Transition / neuron projection arborization / manchette / flagellated sperm motility / cerebellar cortex morphogenesis / positive regulation of cilium assembly / dentate gyrus development / MHC class II antigen presentation / cell projection organization / negative regulation of JNK cascade / UTP biosynthetic process / CTP biosynthetic process / determination of left/right symmetry / centrosome cycle / microtubule organizing center / pyramidal neuron differentiation / motor behavior / intermediate filament / GTP biosynthetic process / nucleoside diphosphate kinase activity / response to L-glutamate / ciliary base / regulation of focal adhesion assembly / negative regulation of T cell receptor signaling pathway / positive regulation of cell motility / tubulin complex / smoothened signaling pathway / intercellular bridge / peptidyl-tyrosine dephosphorylation involved in inactivation of protein kinase activity / regulation of synapse organization Similarity search - Function | ||||||||||||
Biological species | Mus musculus (house mouse) | ||||||||||||
Method | subtomogram averaging / cryo EM / Resolution: 7.7 Å | ||||||||||||
Authors | Chen Z / Shiozak M / Hass KM / Skinner W / Zhao S / Guo C / Polacco BJ / Yu Z / Krogan NJ / Kaake RM ...Chen Z / Shiozak M / Hass KM / Skinner W / Zhao S / Guo C / Polacco BJ / Yu Z / Krogan NJ / Kaake RM / Vale RD / Agard DA | ||||||||||||
Funding support | United States, 3 items
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Citation | Journal: Cell / Year: 2023 Title: De novo protein identification in mammalian sperm using in situ cryoelectron tomography and AlphaFold2 docking. Authors: Zhen Chen / Momoko Shiozaki / Kelsey M Haas / Will M Skinner / Shumei Zhao / Caiying Guo / Benjamin J Polacco / Zhiheng Yu / Nevan J Krogan / Polina V Lishko / Robyn M Kaake / Ronald D Vale / David A Agard / Abstract: To understand the molecular mechanisms of cellular pathways, contemporary workflows typically require multiple techniques to identify proteins, track their localization, and determine their ...To understand the molecular mechanisms of cellular pathways, contemporary workflows typically require multiple techniques to identify proteins, track their localization, and determine their structures in vitro. Here, we combined cellular cryoelectron tomography (cryo-ET) and AlphaFold2 modeling to address these questions and understand how mammalian sperm are built in situ. Our cellular cryo-ET and subtomogram averaging provided 6.0-Å reconstructions of axonemal microtubule structures. The well-resolved tertiary structures allowed us to unbiasedly match sperm-specific densities with 21,615 AlphaFold2-predicted protein models of the mouse proteome. We identified Tektin 5, CCDC105, and SPACA9 as novel microtubule-associated proteins. These proteins form an extensive interaction network crosslinking the lumen of axonemal doublet microtubules, suggesting their roles in modulating the mechanical properties of the filaments. Indeed, Tekt5 -/- sperm possess more deformed flagella with 180° bends. Together, our studies presented a cellular visual proteomics workflow and shed light on the in vivo functions of Tektin 5. | ||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_41431.map.gz | 20.4 MB | EMDB map data format | |
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Header (meta data) | emd-41431-v30.xml emd-41431.xml | 54.2 KB 54.2 KB | Display Display | EMDB header |
Images | emd_41431.png | 136 KB | ||
Filedesc metadata | emd-41431.cif.gz | 16.2 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-41431 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-41431 | HTTPS FTP |
-Validation report
Summary document | emd_41431_validation.pdf.gz | 566.8 KB | Display | EMDB validaton report |
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Full document | emd_41431_full_validation.pdf.gz | 566.4 KB | Display | |
Data in XML | emd_41431_validation.xml.gz | 6.3 KB | Display | |
Data in CIF | emd_41431_validation.cif.gz | 7.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-41431 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-41431 | HTTPS FTP |
-Related structure data
Related structure data | 8to0MC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_41431.map.gz / Format: CCP4 / Size: 80.2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Final map of 48 nm-repeating unit of mouse sperm doublets | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.65 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Sample components
+Entire : Mouse sperm
+Supramolecule #1: Mouse sperm
+Macromolecule #1: Cilia- and flagella-associated protein 95
+Macromolecule #2: Tektin-1
+Macromolecule #3: Detyrosinated tubulin alpha-1A chain
+Macromolecule #4: Tubulin beta-4B chain
+Macromolecule #5: Tektin-2
+Macromolecule #6: EF-hand domain-containing family member B
+Macromolecule #7: Tektin-3
+Macromolecule #8: Meiosis-specific nuclear structural protein 1
+Macromolecule #9: Tektin-4
+Macromolecule #10: Cilia- and flagella-associated protein 53
+Macromolecule #11: Tektin bundle-interacting protein 1
+Macromolecule #12: Cilia- and flagella-associated protein 141
+Macromolecule #13: Nucleoside diphosphate kinase 7
+Macromolecule #14: Protein FAM166B
+Macromolecule #15: Sperm-associated antigen 8
+Macromolecule #16: RIB43A-like with coiled-coils protein 2
+Macromolecule #17: Cilia- and flagella-associated protein 107
+Macromolecule #18: Cilia- and flagella-associated protein 161
+Macromolecule #19: EF-hand domain-containing protein 1
+Macromolecule #20: Sperm acrosome-associated protein 9
+Macromolecule #21: EF-hand domain-containing family member C2
+Macromolecule #22: Cilia- and flagella-associated protein 20
+Macromolecule #23: Parkin coregulated gene protein homolog
+Macromolecule #24: Dual specificity protein phosphatase 3
+Macromolecule #25: Piercer of microtubule wall 2 protein
+Macromolecule #26: Tektin-5
+Macromolecule #27: Cilia- and flagella-associated protein 45
+Macromolecule #28: Cilia- and flagella-associated protein 52
+Macromolecule #29: Enkurin
+Macromolecule #30: Protein Flattop
+Macromolecule #31: Cilia- and flagella- associated protein 210
+Macromolecule #32: Cilia- and flagella-associated protein 276
+Macromolecule #33: EF-hand calcium-binding domain-containing protein 6
+Macromolecule #34: Coiled-coil domain-containing protein 105
+Macromolecule #35: Piercer of microtubule wall 1 protein
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | cell |
-Sample preparation
Buffer | pH: 7.4 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 4.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 6.0 µm / Nominal defocus min: 2.0 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 7.7 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 4.0-beta2) / Number subtomograms used: 12848 |
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Extraction | Number tomograms: 76 / Number images used: 32288 Details: 32288 particles were initially picked every 24 nm along the microtubules. |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |