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-Structure paper
タイトル | DNA origami signposts for identifying proteins on cell membranes by electron cryotomography. |
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ジャーナル・号・ページ | Cell, Vol. 184, Issue 4, Page 1110-1121.e16, Year 2021 |
掲載日 | 2021年2月18日 |
著者 | Emma Silvester / Benjamin Vollmer / Vojtěch Pražák / Daven Vasishtan / Emily A Machala / Catheryne Whittle / Susan Black / Jonathan Bath / Andrew J Turberfield / Kay Grünewald / Lindsay A Baker / |
PubMed 要旨 | Electron cryotomography (cryoET), an electron cryomicroscopy (cryoEM) modality, has changed our understanding of biological function by revealing the native molecular details of membranes, viruses, ...Electron cryotomography (cryoET), an electron cryomicroscopy (cryoEM) modality, has changed our understanding of biological function by revealing the native molecular details of membranes, viruses, and cells. However, identification of individual molecules within tomograms from cryoET is challenging because of sample crowding and low signal-to-noise ratios. Here, we present a tagging strategy for cryoET that precisely identifies individual protein complexes in tomograms without relying on metal clusters. Our method makes use of DNA origami to produce "molecular signposts" that target molecules of interest, here via fluorescent fusion proteins, providing a platform generally applicable to biological surfaces. We demonstrate the specificity of signpost origami tags (SPOTs) in vitro as well as their suitability for cryoET of membrane vesicles, enveloped viruses, and the exterior of intact mammalian cells. |
リンク | Cell / PubMed:33606980 / PubMed Central |
手法 | EM (サブトモグラム平均) |
解像度 | 36.44 Å |
構造データ | EMDB-12188: Structure of DNA origami signpost from cryo-ET and subvolume averaging |
由来 |
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キーワード | DNA / Origami / Nanostructure / Signpost / SPOT |