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TitleA rapid, simple, and economical method for the isolation of ribosomes and translational machinery for structural and functional studies.
Journal, issue, pagesNat Commun, Vol. 16, Issue 1, Page 7185, Year 2025
Publish dateAug 5, 2025
AuthorsJessey Erath / Danielle Kemper / Elisha Mugo / Alex Jacoby / Elizabeth Valenzuela / Courtney F Jungers / Wandy L Beatty / Yaser Hashem / Marko Jovanovic / Sergej Djuranovic / Slavica Pavlovic Djuranovic /
PubMed AbstractRibosomes are RNA-protein complexes essential for protein synthesis and quality control. Traditional methods for ribosome isolation are labor-intensive, expensive, and require a substantial amount of ...Ribosomes are RNA-protein complexes essential for protein synthesis and quality control. Traditional methods for ribosome isolation are labor-intensive, expensive, and require a substantial amount of biological material. In contrast, our method, RNA affinity purification using poly-lysine (RAPPL), provides a rapid, simple, and cost-effective alternative applicable to various species and types of starting material (cell lysates, whole cells, organs, or whole organisms). It is also compatible with traditional isolation techniques. Here, we describe the use of RAPPL for rapid isolation, functional screening, and structural analysis of ribosomes and associated factors. We also demonstrate the application of RAPPL in investigating ribosome-associated resistance mechanisms in uropathogenic Escherichia coli samples and generating a 2.7-Å cryoEM ribosome structure from Cryptococcus neoformans. By significantly reducing the amount of the starting biological material and the time required for isolation, RAPPL has the potential to facilitate the study of ribosomal function, interactions, and antibiotic resistance and provide a versatile platform for academic, clinical, and industrial research.
External linksNat Commun / PubMed:40764614 / PubMed Central
MethodsEM (single particle)
Resolution2.7 Å
Structure data

EMDB-47217: Cryptococcus neoformans 40S Map
Method: EM (single particle) / Resolution: 2.7 Å

EMDB-47218: Cryptococcus neoformans 60S ribosome
Method: EM (single particle) / Resolution: 2.7 Å

EMDB-47219: Cryptococcus neoformans 40S Head Map
Method: EM (single particle) / Resolution: 2.7 Å

Source
  • Cryptococcus neoformans (Cryptococcus neoformans serotype A)

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