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TitleDNA origami signposts for identifying proteins on cell membranes by electron cryotomography.
Journal, issue, pagesCell, Vol. 184, Issue 4, Page 1110-1121.e16, Year 2021
Publish dateFeb 18, 2021
AuthorsEmma 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 AbstractElectron 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.
External linksCell / PubMed:33606980 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution36.44 Å
Structure data

12188

7bho

EMDB-12188: Structure of DNA origami signpost from cryo-ET and subvolume averaging
PDB-7bho: DNA origami signpost designed model
Method: EM (subtomogram averaging) / Resolution: 36.44 Å

Source
  • escherichia virus m13
KeywordsDNA / Origami / Nanostructure / Signpost / SPOT

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