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Yorodumi- EMDB-41431: 48-nm repeating structure of doublets from mouse sperm flagella -
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Open data
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Basic information
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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 | ||||||||||||
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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 informationERKs are inactivated / receptor signaling protein tyrosine kinase inhibitor activity / protein localization to motile cilium / epithelial cilium movement involved in determination of left/right asymmetry / left/right pattern formation / outer acrosomal membrane / regulation of brood size / establishment of left/right asymmetry / 9+0 motile cilium / axonemal microtubule doublet inner sheath ...ERKs are inactivated / receptor signaling protein tyrosine kinase inhibitor activity / protein localization to motile cilium / epithelial cilium movement involved in determination of left/right asymmetry / left/right pattern formation / outer acrosomal membrane / regulation of brood size / establishment of left/right asymmetry / 9+0 motile cilium / axonemal microtubule doublet inner sheath / sperm flagellum assembly / manchette assembly / axonemal B tubule inner sheath / axonemal A tubule inner sheath / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cargo trafficking to the periciliary membrane / Sealing of the nuclear envelope (NE) by ESCRT-III / sperm axoneme assembly / protein polyglutamylation / MAP kinase phosphatase activity / positive regulation of feeding behavior / regulation of calcineurin-NFAT signaling cascade / Intraflagellar transport / Carboxyterminal post-translational modifications of tubulin / regulation of microtubule nucleation / cerebrospinal fluid circulation / inner dynein arm assembly / COPI-independent Golgi-to-ER retrograde traffic / epithelial cilium movement involved in extracellular fluid movement / cilium-dependent cell motility / protein tyrosine/serine/threonine phosphatase activity / regulation of cilium beat frequency involved in ciliary motility / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / 9+2 motile cilium / negative regulation of chemotaxis / left/right axis specification / regulation of store-operated calcium entry / COPI-mediated anterograde transport / cilium movement involved in cell motility / intraciliary transport / Kinesins / acrosomal membrane / axoneme assembly / cilium movement / PKR-mediated signaling / Aggrephagy / microtubule sliding / RHO GTPases activate IQGAPs / COPI-dependent Golgi-to-ER retrograde traffic / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / Resolution of Sister Chromatid Cohesion / motile cilium assembly / perinuclear theca / Recycling pathway of L1 / The role of GTSE1 in G2/M progression after G2 checkpoint / calcium ion sensor activity / axonemal microtubule / Hedgehog 'off' state / RHO GTPases Activate Formins / cilium organization / Loss of Nlp from mitotic centrosomes / Recruitment of mitotic centrosome proteins and complexes / Loss of proteins required for interphase microtubule organization from the centrosome / organelle transport along microtubule / Anchoring of the basal body to the plasma membrane / Separation of Sister Chromatids / Recruitment of NuMA to mitotic centrosomes / AURKA Activation by TPX2 / gamma-tubulin ring complex / forebrain morphogenesis / glial cell differentiation / cellular response to leukemia inhibitory factor / cerebellar cortex morphogenesis / dentate gyrus development / manchette / determination of left/right symmetry / Regulation of PLK1 Activity at G2/M Transition / positive regulation of cilium assembly / positive regulation of focal adhesion disassembly / neuron projection arborization / MHC class II antigen presentation / 3'-5'-DNA exonuclease activity / smoothened signaling pathway / UTP biosynthetic process / CTP biosynthetic process / motor behavior / pyramidal neuron differentiation / sperm head-tail coupling apparatus / motile cilium / DNA catabolic process / negative regulation of JNK cascade / dephosphorylation / response to L-glutamate / centrosome cycle / nucleoside diphosphate kinase activity / single fertilization / mRNA stabilization / positive regulation of cell motility / regulation of focal adhesion assembly Similarity search - Function | ||||||||||||
| Biological species | ![]() | ||||||||||||
| 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: 2023Title: 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 |
-Related structure data
| Related structure data | ![]() 8to0MC C: citing same article ( M: atomic model generated by this map |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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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
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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 |
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Sample preparation
| Buffer | pH: 7.4 |
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| Vitrification | Cryogen name: ETHANE |
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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 |
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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 |
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Keywords
Authors
United States, 3 items
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FIELD EMISSION GUN
