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EMD-41431

Subtomogram averaging
7.7 Å
EMD-41431 Deposition: 02/08/2023
Map released: 01/11/2023
Last modified: 22/11/2023
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links

EMD-41431

48-nm repeating structure of doublets from mouse sperm flagella

EMD-41431

Subtomogram averaging
7.7 Å
EMD-41431 Deposition: 02/08/2023
Map released: 01/11/2023
Last modified: 22/11/2023
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Sample Organism: Mus musculus
Sample: Mouse sperm
Fitted models: 8to0

Deposition Authors: Chen Z , Shiozak M, Hass KM, Skinner W, Zhao S, Guo C, Polacco BJ, Yu Z, Krogan NJ, Kaake RM, Vale RD, Agard DA
De novo protein identification in mammalian sperm using in situ cryoelectron tomography and AlphaFold2 docking.
PUBMED: 37865089
DOI: doi:10.1016/j.cell.2023.09.017
ISSN: 1097-4172
Abstract:
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.