Abstract
Current research is focusing on ribosome heterogeneity as a response to changing environmental conditions and stresses. Altered stoichiometry and composition of ribosomal proteins as well as association of additional protein factors are mechanisms for shaping the protein expression profile or hibernating ribosomes. In this updated chapter, we present a method for the isolation of ribosomes to analyze antibiotic-induced changes in the composition of ribosomes in Bacillus subtilis or other bacteria. Ribosomes and associated proteins are isolated by ultracentrifugation, and proteins are identified and quantified using label-free mass spectrometry.
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Acknowledgments
We thank Birgit Klinkert and Johanna Roßmanith for the practical introduction into ribosome isolation and for technical support. Furthermore, we would like to thank Dörte Becher and Knut Büttner for sharing mass spectrometry protocols. Funding from the German Federal State of North Rhine Westphalia (NRW) is acknowledged for the mass spectrometer (“Forschungsgroßgeräte der Länder”) used in this protocol. JEB acknowledges funding from NRW from the grant “Translation of innovative antibiotics from NRW” for performing the original work and preparing the original chapter and from NRW and the European Union, European Regional Development Fund, Investing in your future (Research Infrastructure “Center for System-based Antibiotic Research (CESAR)”) for updating the chapter.
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Schäkermann, S., Dietze, P., Bandow, J.E. (2023). Label-Free Quantitation of Ribosomal Proteins from Bacillus subtilis for Antibiotic Research. In: Sass, P. (eds) Antibiotics. Methods in Molecular Biology, vol 2601. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2855-3_20
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DOI: https://doi.org/10.1007/978-1-0716-2855-3_20
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