A rapid, simple, and economical method for the isolation of ribosomes and translational machinery for structural and functional studies

Jessey Erath, Danielle Kemper, Elisha Mugo, Alex Jacoby, Elizabeth Valenzuela, Courtney F. Jungers, Wandy L. Beatty, Yaser Hashem, Marko Jovanovic, Sergej Djuranovic () and Slavica Pavlovic Djuranovic ()
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Jessey Erath: Washington University School of Medicine
Danielle Kemper: Washington University School of Medicine
Elisha Mugo: Washington University School of Medicine
Alex Jacoby: Washington University School of Medicine
Elizabeth Valenzuela: Columbia University
Courtney F. Jungers: Washington University School of Medicine
Wandy L. Beatty: Washington University School of Medicine
Yaser Hashem: Université de Bordeaux
Marko Jovanovic: Columbia University
Sergej Djuranovic: Washington University School of Medicine
Slavica Pavlovic Djuranovic: Washington University School of Medicine

Nature Communications, 2025, vol. 16, issue 1, 1-16

Abstract: Abstract 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.

Date: 2025
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-62314-8

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DOI: 10.1038/s41467-025-62314-8

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