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| Verfasst von: | Fung, Herman K. H. [VerfasserIn]  |
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| | Hayashi, Yuki [VerfasserIn] |
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| | Salo, Veijo T. [VerfasserIn] |
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| | Babenko, Anastasiia [VerfasserIn] |
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| | Zagoriy, Ievgeniia [VerfasserIn] |
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| | Brunner, Andreas [VerfasserIn] |
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| | Ellenberg, Jan [VerfasserIn] |
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| | Müller, Christoph W. [VerfasserIn] |
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| | Cuylen-Haering, Sara [VerfasserIn] |
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| | Mahamid, Julia [VerfasserIn] |
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| Titel: | Genetically encoded multimeric tags for subcellular protein localization in cryo-EM |
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| Verf.angabe: | Herman K.H. Fung, Yuki Hayashi, Veijo T. Salo, Anastasiia Babenko, Ievgeniia Zagoriy, Andreas Brunner, Jan Ellenberg, Christoph W. Müller, Sara Cuylen-Haering & Julia Mahamid |
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| E-Jahr: | 2023 |
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| Jahr: | 06 November 2023 |
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| Umfang: | 30 S. |
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| Fussnoten: | Gesehen am 03.06.2023 |
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| Titel Quelle: | Enthalten in: Nature methods |
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| Ort Quelle: | London [u.a.] : Nature Publishing Group, 2004 |
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| Jahr Quelle: | 2023 |
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| Band/Heft Quelle: | 20(2023), 12, Seite 1900-1929 |
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| ISSN Quelle: | 1548-7105 |
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| Abstract: | Abstract - Cryo-electron tomography (cryo-ET) allows for label-free high-resolution imaging of macromolecular assemblies in their native cellular context. However, the localization of macromolecules of interest in tomographic volumes can be challenging. Here we present a ligand-inducible labeling strategy for intracellular proteins based on fluorescent, 25-nm-sized, genetically encoded multimeric particles (GEMs). The particles exhibit recognizable structural signatures, enabling their automated detection in cryo-ET data by convolutional neural networks. The coupling of GEMs to green fluorescent protein-tagged macromolecules of interest is triggered by addition of a small-molecule ligand, allowing for time-controlled labeling to minimize disturbance to native protein function. We demonstrate the applicability of GEMs for subcellular-level localization of endogenous and overexpressed proteins across different organelles in human cells using cryo-correlative fluorescence and cryo-ET imaging. We describe means for quantifying labeling specificity and efficiency, and for systematic optimization for rare and abundant protein targets, with emphasis on assessing the potential effects of labeling on protein function. |
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| DOI: | doi:10.1038/s41592-023-02053-0 |
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| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
kostenfrei: Volltext: https://doi.org/10.1038/s41592-023-02053-0 |
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| | kostenfrei: Volltext: https://www.nature.com/articles/s41592-023-02053-0 |
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| | DOI: https://doi.org/10.1038/s41592-023-02053-0 |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| K10plus-PPN: | 1890494976 |
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| Verknüpfungen: | → Zeitschrift |
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Genetically encoded multimeric tags for subcellular protein localization in cryo-EM / Fung, Herman K. H. [VerfasserIn]; 06 November 2023 (Online-Ressource)
