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Phosphoproteomic Analysis of Isolated Mitochondria in Yeast

  • Margaux Renvoisé
  • Ludovic Bonhomme
  • Marlène Davanture
  • Michel Zivy
  • Claire LemaireEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1636)

Abstract

Mitochondria play a central role in cellular energy metabolism and cell death. Deregulation of mitochondrial functions is associated with several human pathologies (neurodegenerative diseases, neuromuscular diseases, type II diabetes, obesity, cancer). The steadily increasing number of identified mitochondrial phosphoproteins, kinases, and phosphatases in recent years suggests that reversible protein phosphorylation plays an important part in the control of mitochondrial processes. In addition, many mitochondrial phosphoproteins probably still remain to be identified, considering that 30% of proteins are expected to be phosphorylated in eukaryotes. In this chapter, we describe two procedures for the analysis of the mitochondrial phosphoproteome. The first one is a qualitative method that combines blue native and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (2D-BN/SDS-PAGE) and specific phosphoprotein staining. The second one is a quantitative approach that associates mitochondrial peptide labeling, phosphopeptide enrichment, and mass spectrometry.

Key words

Saccharomyces cerevisiae Mitochondria Phosphoproteome 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Margaux Renvoisé
    • 1
  • Ludovic Bonhomme
    • 2
  • Marlène Davanture
    • 3
  • Michel Zivy
    • 3
  • Claire Lemaire
    • 1
    Email author
  1. 1.UMR 9198 CNRS, Institute for Integrative Biology of the Cell (I2BC), B3S, LPSM—CEA SaclayGif-sur-Yvette cedexFrance
  2. 2.INRA/UCA UMR 1095 GDEC ‘Génétique, Diversité et Ecophysiologie des Céréales’Clermont-FerrandFrance
  3. 3.GQE– Le Moulon, INRA, Univ. Paris-Sud, CNRS, AgroParisTech, UniversitÕ Paris-SaclayGif-sur-YvetteFrance

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