Yeast Mitoribosome Purification and Analyses by Sucrose Density Centrifugation and Immunoprecipitation

Aufschnaiter, Andreas,Dr. rer. nat.1988-Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden (author)

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.

Carlström, AndreasDepartment of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden (author)
Ott, MartinDepartment of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden; Department of Medical Biochemistry and Cell Biology, University of Gothenburg, Gothenburg, Sweden (author)
Department of Biochemistry and Biophysics, Stockholm University, Stockholm, SwedenDepartment of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden; Department of Medical Biochemistry and Cell Biology, University of Gothenburg, Gothenburg, Sweden (creator_code:org_t)

By the author/editor: Aufschnaiter, An ...; Carlström, Andre ...; Ott, Martin

About the subject

NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...; and Biochemistry and ...

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