Chapter title |
Mass Spectrometry of Mitochondrial Membrane Protein Complexes
|
---|---|
Chapter number | 12 |
Book title |
Membrane Protein Structure and Function Characterization
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7151-0_12 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7149-7, 978-1-4939-7151-0
|
Authors |
Luc Negroni, Michel Zivy, Claire Lemaire |
Abstract |
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 (1) permits the separation of functional respiratory complexes, thus allowing in-gel activity detection of most of the respiratory complexes and (2) 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 of highlighting potential interacting proteins. |
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Geographical breakdown
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Unknown | 13 | 100% |
Demographic breakdown
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Researcher | 5 | 38% |
Student > Ph. D. Student | 2 | 15% |
Student > Doctoral Student | 1 | 8% |
Unspecified | 1 | 8% |
Student > Master | 1 | 8% |
Other | 1 | 8% |
Unknown | 2 | 15% |
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Unspecified | 1 | 8% |
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Neuroscience | 1 | 8% |
Other | 1 | 8% |
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