TY - JOUR T1 - Monitoring mitochondrial translation by pulse SILAC JF - bioRxiv DO - 10.1101/2021.01.31.428997 SP - 2021.01.31.428997 AU - Koshi Imami AU - Matthias Selbach AU - Yasushi Ishihama Y1 - 2021/01/01 UR - http://biorxiv.org/content/early/2021/02/01/2021.01.31.428997.abstract N2 - Mitochondrial ribosomes are specialized to translate the 13 membrane proteins encoded in the mitochondrial genome, but it is challenging to quantify mitochondrial translation products due to their hydrophobic nature. Here, we introduce a proteomic method that combines biochemical isolation of mitochondria with pulse stable isotope labeling by amino acids in cell culture (pSILAC). Our method provides the highest protein coverage (quantifying 12 out of the 13 inner-membrane proteins; average 2-fold improvement over previous studies) with the shortest measurement time. We applied this method to uncover the global picture of (post)translational regulation of both mitochondrial- and nuclear-encoded proteins involved in the assembly of protein complexes that mediate oxidative phosphorylation (OXPHOS). The results allow us to infer the assembly order of complex components and/or partners, as exemplified by complex III. This method should be applicable to study mitochondrial translation programs in many contexts, including oxidative stress and mitochondrial disease.Competing Interest StatementThe authors have declared no competing interest. ER -