title: Identification of lysine-acetylated mitochondrial proteins and their acetylation sites.
abstract: The (ε)N-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.
authors.affiliation: Plant Proteomics and Mass, Spectrometry Group, Max-Planck-Institute for Plant Breeding Research, Carl-von-Linné Weg 10, 50829, Cologne, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Hartl
authors.forename: Ann-Christine
authors.initials: AC
authors.lastname: König
authors.forename: Iris
authors.initials: I
authors.lastname: Finkemeier
mesh_terms: D000107:Acetylation; D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins / Q000737:chemistry*; D015153:Blotting, Western / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D008239:Lysine / Q000032:analysis*; D008928:Mitochondria / Q000737:chemistry; D024101:Mitochondrial Proteins / Q000737:chemistry*; D063347:Native Polyacrylamide Gel Electrophoresis / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D029681:Arabidopsis Proteins; D024101:Mitochondrial Proteins; D008239:Lysine