Cryo-electron tomography pipeline for plasma membranes
Abstract Cryo-electron tomography (cryoET) provides sub-nanometer protein structure within the dense cellular environment. Existing sample preparation methods are insufficient at accessing the plasma membrane and its associated proteins. Here, we present a correlative cryo-electron tomography pipeli...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56045-z |
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| author | Willy W. Sun Dennis J. Michalak Kem A. Sochacki Prasanthi Kunamaneni Marco A. Alfonzo-Méndez Andreas M. Arnold Marie-Paule Strub Jenny E. Hinshaw Justin W. Taraska |
| author_facet | Willy W. Sun Dennis J. Michalak Kem A. Sochacki Prasanthi Kunamaneni Marco A. Alfonzo-Méndez Andreas M. Arnold Marie-Paule Strub Jenny E. Hinshaw Justin W. Taraska |
| author_sort | Willy W. Sun |
| collection | DOAJ |
| description | Abstract Cryo-electron tomography (cryoET) provides sub-nanometer protein structure within the dense cellular environment. Existing sample preparation methods are insufficient at accessing the plasma membrane and its associated proteins. Here, we present a correlative cryo-electron tomography pipeline optimally suited to image large ultra-thin areas of isolated basal and apical plasma membranes. The pipeline allows for angstrom-scale structure determination with subtomogram averaging and employs a genetically encodable rapid chemically-induced electron microscopy visible tag for marking specific proteins within the complex cellular environment. The pipeline provides efficient, distributable, low-cost sample preparation and enables targeted structural studies of identified proteins at the plasma membrane of mammalian cells. |
| format | Article |
| id | doaj-art-27c7c06964774b0b83eeafad22e465fa |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-27c7c06964774b0b83eeafad22e465fa2025-02-09T12:43:55ZengNature PortfolioNature Communications2041-17232025-01-0116111410.1038/s41467-025-56045-zCryo-electron tomography pipeline for plasma membranesWilly W. Sun0Dennis J. Michalak1Kem A. Sochacki2Prasanthi Kunamaneni3Marco A. Alfonzo-Méndez4Andreas M. Arnold5Marie-Paule Strub6Jenny E. Hinshaw7Justin W. Taraska8National Heart, Lung, and Blood Institute, US National Institutes of HealthNational Heart, Lung, and Blood Institute, US National Institutes of HealthNational Heart, Lung, and Blood Institute, US National Institutes of HealthNational Heart, Lung, and Blood Institute, US National Institutes of HealthNational Heart, Lung, and Blood Institute, US National Institutes of HealthNational Heart, Lung, and Blood Institute, US National Institutes of HealthNational Heart, Lung, and Blood Institute, US National Institutes of HealthNational Institute of Diabetes and Digestive and Kidney Diseases, US National Institutes of HealthNational Heart, Lung, and Blood Institute, US National Institutes of HealthAbstract Cryo-electron tomography (cryoET) provides sub-nanometer protein structure within the dense cellular environment. Existing sample preparation methods are insufficient at accessing the plasma membrane and its associated proteins. Here, we present a correlative cryo-electron tomography pipeline optimally suited to image large ultra-thin areas of isolated basal and apical plasma membranes. The pipeline allows for angstrom-scale structure determination with subtomogram averaging and employs a genetically encodable rapid chemically-induced electron microscopy visible tag for marking specific proteins within the complex cellular environment. The pipeline provides efficient, distributable, low-cost sample preparation and enables targeted structural studies of identified proteins at the plasma membrane of mammalian cells.https://doi.org/10.1038/s41467-025-56045-z |
| spellingShingle | Willy W. Sun Dennis J. Michalak Kem A. Sochacki Prasanthi Kunamaneni Marco A. Alfonzo-Méndez Andreas M. Arnold Marie-Paule Strub Jenny E. Hinshaw Justin W. Taraska Cryo-electron tomography pipeline for plasma membranes Nature Communications |
| title | Cryo-electron tomography pipeline for plasma membranes |
| title_full | Cryo-electron tomography pipeline for plasma membranes |
| title_fullStr | Cryo-electron tomography pipeline for plasma membranes |
| title_full_unstemmed | Cryo-electron tomography pipeline for plasma membranes |
| title_short | Cryo-electron tomography pipeline for plasma membranes |
| title_sort | cryo electron tomography pipeline for plasma membranes |
| url | https://doi.org/10.1038/s41467-025-56045-z |
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