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TitleRapid structural analysis of bacterial ribosomes in situ.
Journal, issue, pagesCommun Biol, Vol. 8, Issue 1, Page 131, Year 2025
Publish dateJan 28, 2025
AuthorsBarrett M Powell / Tyler S Brant / Joseph H Davis / Shyamal Mosalaganti /
PubMed AbstractRapid structural analysis of purified proteins and their complexes has become increasingly common thanks to key methodological advances in cryo-electron microscopy (cryo-EM) and associated data ...Rapid structural analysis of purified proteins and their complexes has become increasingly common thanks to key methodological advances in cryo-electron microscopy (cryo-EM) and associated data processing software packages. In contrast, analogous structural analysis in cells via cryo-electron tomography (cryo-ET) remains challenging due to critical technical bottlenecks, including low-throughput sample preparation and imaging, and laborious data processing methods. Here, we describe a rapid in situ cryo-ET sample preparation and data analysis workflow that results in the routine determination of sub-nm resolution ribosomal structures. We apply this workflow to E. coli, producing a 5.8 Å structure of the 70S ribosome from cells in less than 10 days and facilitating the discovery of a minor population of 100S-like disomes. We envision our approach to be widely applicable to related bacterial samples.
External linksCommun Biol / PubMed:39875527 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution5.88 - 15.0 Å
Structure data

EMDB-44152: E.coli 70S ribosome imaged in situ with rapid processing pipeline
Method: EM (subtomogram averaging) / Resolution: 5.88 Å

EMDB-45638: Non-canonical E.coli 100S ribosome imaged in situ with rapid processing pipeline
Method: EM (subtomogram averaging) / Resolution: 15.0 Å

Source
  • Escherichia coli (E. coli)

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