Yeast Mitoribosome Purification and Analyses by Sucrose Density Centrifugation and Immunoprecipitation

Andreas Aufschnaiter; Andreas Carlström; Martin Ott

doi:10.1007/978-1-0716-3171-3_8

Yeast Mitoribosome Purification and Analyses by Sucrose Density Centrifugation and Immunoprecipitation

Methods Mol Biol. 2023:2661:119-132. doi: 10.1007/978-1-0716-3171-3_8.

Authors

Andreas Aufschnaiter^#¹, Andreas Carlström^#¹, Martin Ott^{2

3}

Affiliations

¹ Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden.
² Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden. martin.ott@gu.se.
³ Department of Medical Biochemistry and Cell Biology, University of Gothenburg, Gothenburg, Sweden. martin.ott@gu.se.

^# Contributed equally.

PMID: 37166635
DOI: 10.1007/978-1-0716-3171-3_8

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.

Keywords: Immunoprecipitation; Mitochondria; Mitoribosome; Sucrose cushion; Sucrose gradient; Yeast.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Immunoprecipitation
Mitochondria / metabolism
Mitochondrial Proteins / metabolism
Mitochondrial Ribosomes* / metabolism
Protein Biosynthesis
Saccharomyces cerevisiae* / genetics

Substances

Mitochondrial Proteins