Abstract
Mitochondrial protein biosynthesis is maintained by an interplay between the mitochondrial ribosome (mitoribosome) and a large set of protein interaction partners. This interactome regulates a diverse set of functions, including mitochondrial gene expression, translation, protein quality control, and respiratory chain assembly. Hence, robust methods to biochemically and structurally analyze this molecular machinery are required to understand the sophisticated regulation of mitochondrial protein biosynthesis. In this chapter, we present detailed protocols for immunoprecipitation, sucrose cushions, and linear sucrose gradients to purify and analyze mitoribosomes and their interaction partners.
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Acknowledgment
This work was supported by the Swedish Research Council and the Knut and Alice Wallenberg foundation (to Martin Ott) and the Austrian Science Fund FWF (J4398-B to Andreas Aufschnaiter).
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Aufschnaiter, A., Carlström, A., Ott, M. (2023). Yeast Mitoribosome Purification and Analyses by Sucrose Density Centrifugation and Immunoprecipitation. In: Barrientos, A., Fontanesi, F. (eds) The Mitoribosome. Methods in Molecular Biology, vol 2661. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3171-3_8
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DOI: https://doi.org/10.1007/978-1-0716-3171-3_8
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