National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01GM136823
United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R01GM112824
United States
Citation
Journal: Nat Commun / Year: 2025 Title: A rapid, simple, and economical method for the isolation of ribosomes and translational machinery for structural and functional studies. Authors: Jessey Erath / Danielle Kemper / Elisha Mugo / Alex Jacoby / Elizabeth Valenzuela / Courtney F Jungers / Wandy L Beatty / Yaser Hashem / Marko Jovanovic / Sergej Djuranovic / Slavica Pavlovic Djuranovic / 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 ...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.
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