Identification of Lysine-Acetylated Mitochondrial Proteins and Their Acetylation Sites

  • Markus Hartl
  • Ann-Christine König
  • Iris Finkemeier
Part of the Methods in Molecular Biology book series (MIMB, volume 1305)

Abstract

The εN-acetylation of lysine side chains is a highly conserved posttranslational modification of both prokaryotic and eukaryotic proteins. Lysine acetylation not only occurs on histones in the nucleus but also on many mitochondrial proteins in plants and animals. As the transfer of the acetyl group to lysine eliminates its positive charge, lysine acetylation can affect the biological function of proteins. This chapter describes two methods for the identification of lysine-acetylated proteins in plant mitochondria using an anti-acetyllysine antibody. We describe the Western blot analysis of a two-dimensional blue native-polyacrylamide gel electrophoresis with an anti-acetyllysine antibody as well as the immuno-enrichment of lysine-acetylated peptides followed by liquid chromatography-tandem mass spectrometry data acquisition and analysis.

Key words

Lysine acetylation Mitochondria Arabidopsis Protein complexes Blue native-polyacrylamide gel electrophoresis (BN-PAGE) 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Markus Hartl
    • 1
  • Ann-Christine König
    • 1
  • Iris Finkemeier
    • 1
  1. 1.Plant Proteomics and Mass, Spectrometry GroupMax-Planck-Institute for Plant Breeding ResearchCologneGermany

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