title: SILAC-based quantification of Sirt1-responsive lysine acetylome.
abstract: Stable Isotope Labeling by Amino acids in Cell culture (SILAC) is one of the in vivo metabolic labeling methods widely used for dynamic analysis of protein modifications. Here, we describe a general approach to applying SILAC, in combination with affinity enrichment of acetyllysine peptides and mass spectrometry, to study the dynamic changes of the Lysine acetylome in response to Sirt1. The method should be applicable to quantify changes to other post translational modifications in diverse cellular systems.
authors.affiliation: Ben May Department for Cancer Research, University of Chicago, Chicago, IL, USA.
authors.forename: Yue
authors.initials: Y
authors.lastname: Chen
authors.forename: Gozde
authors.initials: G
authors.lastname: Colak
authors.forename: Yingming
authors.initials: Y
authors.lastname: Zhao
mesh_terms: D000107:Acetylation; D000818:Animals; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015151:Immunoblotting; D047468:Immunoprecipitation / Q000379:methods*; D007553:Isotope Labeling / Q000379:methods*; D008239:Lysine / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D011499:Protein Processing, Post-Translational; D056564:Sirtuin 1 / Q000502:physiology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: C470545:Sirt1 protein, mouse; D056564:Sirtuin 1; D008239:Lysine