7Q23

cryo iDPC-STEM structure recorded with CSA 3.0

Summary for 7Q23

• Entry DOI: 10.2210/pdb7q23/pdb
• EMDB information: 13779
• Descriptor: Capsid protein, RNA (5'-R(P*GP*AP*A)-3') (2 entities in total)
• Functional Keywords: tobacco mosaic virus, rna virus, virus
• Biological source: Tobacco mosaic virus (strain vulgare) (TMV, Tobacco mosaic virus (strain U1)); More
• Total number of polymer chains: 2
• Total formula weight: 18050.66

Authors

Sachse, C.,Leidl, M.L. (deposition date: 2021-10-22, release date: 2022-09-21, Last modification date: 2024-07-17)

Primary citation

Lazic, I.,Wirix, M.,Leidl, M.L.,de Haas, F.,Mann, D.,Beckers, M.,Pechnikova, E.V.,Muller-Caspary, K.,Egoavil, R.,Bosch, E.G.T.,Sachse, C.
Single-particle cryo-EM structures from iDPC-STEM at near-atomic resolution.
Nat.Methods, 19:1126-1136, 2022

PubMed Abstract: In electron cryomicroscopy (cryo-EM), molecular images of vitrified biological samples are obtained by conventional transmission microscopy (CTEM) using large underfocuses and subsequently computationally combined into a high-resolution three-dimensional structure. Here, we apply scanning transmission electron microscopy (STEM) using the integrated differential phase contrast mode also known as iDPC-STEM to two cryo-EM test specimens, keyhole limpet hemocyanin (KLH) and tobacco mosaic virus (TMV). The micrographs show complete contrast transfer to high resolution and enable the cryo-EM structure determination for KLH at 6.5 Å resolution, as well as for TMV at 3.5 Å resolution using single-particle reconstruction methods, which share identical features with maps obtained by CTEM of a previously acquired same-sized TMV data set. These data show that STEM imaging in general, and in particular the iDPC-STEM approach, can be applied to vitrified single-particle specimens to determine near-atomic resolution cryo-EM structures of biological macromolecules.

PubMed: 36064775
DOI: 10.1038/s41592-022-01586-0

Experimental method

ELECTRON MICROSCOPY (4.3 Å)