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| Title | Capturing ribosomal structures in cellular extracts with cryoPRISM: A purification-free cryoEM approach reveals novel structural states. |
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| Journal, issue, pages | Proc Natl Acad Sci U S A, Vol. 123, Issue 9, Page e2521210123, Year 2026 |
| Publish date | Mar 3, 2026 |
 Authors | Mira B May / Gabriella S Lopez-Perez / Joseph H Davis /  |
| 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 ...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. |
 External links | Proc Natl Acad Sci U S A / PubMed:41739560 / PubMed Central |
| Methods | EM (single particle) |
| Resolution | 2.57 - 4.0 Å |
| Structure data | EMDB-72030, PDB-9pyc:  EMDB-72250: Cryo-EM structure of 70S ribosome in the classical state (A/A, P/P, E,E) imaged in a cell lysate  EMDB-72251: Cryo-EM structure of 70S ribosome in the classical state (P/P, E/E) imaged in a cell lysate  EMDB-72255: Cryo-EM structure of 70S ribosome bound to RaiA imaged in a cell lysate  EMDB-72257: Cryo-EM structure of a tmRNA-rescue complex imaged in a cell lysate  EMDB-72271: Cryo-EM structure of 70S ribosome bound to EF-Tu imaged in a cell lysate  EMDB-72291: Cryo-EM structure of 30S ribosome in a H44 active state imaged in a cell lysate  EMDB-72292: Cryo-EM structure of 30S ribosome in a H44 inactive-dislodged state imaged in a cell lysate  EMDB-72315: Cryo-EM structure of 30S ribosome in a H44 inactive-unresolved state imaged in a cell lysate  EMDB-72316: Cryo-EM structure of 30S ribosome in a closed conformation with tRNA bound imaged in a cell lysate  EMDB-72324: Cryo-EM structure of 30S ribosome with tRNA bound in an open conformation imaged in a cell lysate  EMDB-72344: Cryo-EM structure of 50S ribosome (class I) imaged in a cell lysate  EMDB-72346: Cryo-EM structure of large ribosomal subunit (class 2) imaged in a cell lysate  EMDB-72349: Cryo-EM structure of large ribosomal subunit (class 3) imaged in a cell lysate  EMDB-72350: Cryo-EM structure of large ribosomal subunit (class 5) imaged in a cell lysate  EMDB-72354: Cryo-EM structure of large ribosomal subunit (class A1) imaged in a cell lysate  EMDB-72355: Cryo-EM structure of large ribosomal subunit (class A2) imaged in a cell lysate  EMDB-72357: Cryo-EM structure of 70S ribosome in the hybrid state (A/P*, P/E) imaged in a cell lysate  EMDB-72360: Cryo-EM structure of large ribosomal subunit (class C1) imaged in a cell lysate  EMDB-72363: Cryo-EM structure of large ribosomal subunit (class B2) imaged in a cell lysate  EMDB-72364: Cryo-EM structure of large ribosomal subunit (class B1) imaged in a cell lysate  EMDB-72365: Cryo-EM structure of large ribosomal subunit (class 4) imaged in a cell lysate  EMDB-72366: Cryo-EM structure of large ribosomal subunit (class 6) imaged in a cell lysate |
| Source |
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 Keywords | RIBOSOMAL PROTEIN / hibernation / elongation factor / bacterial ribosome |
escherichia coli k-12 (bacteria)