9PYC

Cryo-EM structure of EF-G and RaiA simultaneously bound to an E. coli ribosome imaged in a cell lysate

9PYC の概要

• エントリーDOI: 10.2210/pdb9pyc/pdb
• EMDBエントリー: 72030
• 分子名称: Elongation factor G, Ribosome-associated inhibitor A (2 entities in total)
• 機能のキーワード: hibernation, elongation factor, bacterial ribosome, ribosomal protein
• 由来する生物種: Escherichia coli K-12; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 90479.81

構造登録者

May, M.B.,Davis, J.H. (登録日: 2025-08-07, 公開日: 2026-02-18, 最終更新日: 2026-03-11)

主引用文献

May, M.B.,Lopez-Perez, G.S.,Davis, J.H.
Capturing ribosomal structures in cellular extracts with cryoPRISM: A purification-free cryoEM approach reveals novel structural states.
Proc.Natl.Acad.Sci.USA, 123:e2521210123-e2521210123, 2026

PubMed Abstract: Structural analyses of ribosomes by single particle cryogenic electron microscopy (cryoEM) have traditionally relied on purified or reconstituted samples, with particles often trapped in desired states using genetic, pharmacological, or biochemical perturbations. While informative, such in vitro methods often fail to capture the full diversity of structural states and associated protein factors present in cells. In contrast, in situ cryoelectron tomography preserves cellular context but is limited by low throughput and modest resolution. Here, we present cryoPRISM (purification-free ribosome imaging from subcellular mixtures), a rapid ex vivo workflow encompassing cell lysis, vitrification, and image analysis methods for high-resolution analyses of ribosomal structures directly from cell lysates. Applying cryoPRISM in , we resolved more than 20 distinct ribosomal states spanning assembly, translation initiation, elongation, trans-translation, and quiescence, including a novel configuration of EF-G bound to idle ribosomes with the ribosome hibernation factor ribosome-associated inhibitor A. Given its speed, accessibility, and ability to preserve native interactions and structural heterogeneity, we anticipate that cryoPRISM will be broadly applicable for uncovering ribosomal biology across diverse organisms and conditions.

PubMed: 41739560
DOI: 10.1073/pnas.2521210123

実験手法

ELECTRON MICROSCOPY (2.9 Å)