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| Title | Molecular goniometers for single-particle cryo-electron microscopy of DNA-binding proteins. |
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| Journal, issue, pages | Nat Biotechnol, Vol. 39, Issue 3, Page 378-386, Year 2021 |
| Publish date | Oct 19, 2020 |
 Authors | Tural Aksel / Zanlin Yu / Yifan Cheng / Shawn M Douglas /  |
| PubMed Abstract | Correct reconstruction of macromolecular structure by cryo-electron microscopy (cryo-EM) relies on accurate determination of the orientation of single-particle images. For small (<100 kDa) DNA-binding proteins, obtaining particle images with sufficiently asymmetric features to correctly guide alignment is challenging. We apply DNA origami to construct molecular goniometers-instruments that precisely orient objects-and use them to dock a DNA-binding protein on a double-helix stage that has user-programmable tilt and rotation angles. We construct goniometers with 14 different stage configurations to orient and visualize the protein just above the cryo-EM grid surface. Each goniometer has a distinct barcode pattern that we use during particle classification to assign angle priors to the bound protein. We use goniometers to obtain a 6.5-Å structure of BurrH, an 82-kDa DNA-binding protein whose helical pseudosymmetry prevents accurate image orientation using traditional cryo-EM. Our approach should be adaptable to other DNA-binding proteins as well as small proteins fused to DNA-binding domains. |
 External links | Nat Biotechnol / PubMed:33077960 / PubMed Central |
| Methods | EM (single particle) |
| Resolution | 6.5 Å |
| Structure data |  EMDB-21443: |
| Source |
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Paraburkholderia rhizoxinica (bacteria)