Mass Spectrometry of Mitochondrial Membrane Protein Complexes
Résumé
The ATP production (oxidative phosphorylation) involves five complexes embedded in the inner membrane of mitochondria. The yeast Saccharomyces cerevisiae is mainly used as a model for the study of oxidative phosphorylation; mutants are easy to produce and are still viable due to their ability to grow using the fermentation pathway. Here we present a process for analyzing mitochondrial respiratory complexes using native electrophoresis (BN-PAGE) coupled to LC-MS/MS. BN-PAGE (i) permits the separation of functional respiratory complexes, thus allowing in-gel activity detection of most of the respiratory complexes and (ii) provides convenient samples for bottom-up proteomics. Combining BN-PAGE and LC-MS/MS leads to the identification of the subunit composition of membrane complexes and offers the possibility to highlight potential interacting proteins.
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MiMB 2017 - Negroni et al. - Mass spectrometry of mitochondrial membrane protein.pdf (233.32 Ko)
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