Rapid structural analysis of bacterial ribosomes in situ .
Barrett M PowellTyler S BrantJoseph H DavisShyamal MosalagantiPublished in: bioRxiv : the preprint server for biology (2024)
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 the development of 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 we expect this workflow will be widely applicable to related bacterial samples.
Keyphrases
- electron microscopy
- data analysis
- electronic health record
- high resolution
- induced apoptosis
- cell cycle arrest
- molecularly imprinted
- escherichia coli
- loop mediated isothermal amplification
- magnetic resonance
- big data
- endoplasmic reticulum stress
- machine learning
- clinical practice
- photodynamic therapy
- artificial intelligence
- cell death
- deep learning
- cell proliferation
- quantum dots
- fluorescence imaging
- drug induced