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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Acetylation
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Animals
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Cells, Cultured
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Chromatography, Affinity
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Chromatography, Liquid / methods*
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Embryo, Mammalian / cytology
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Embryo, Mammalian / metabolism
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Fibroblasts / cytology
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Fibroblasts / metabolism
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Immunoblotting
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Immunoprecipitation / methods*
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Isotope Labeling / methods*
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Lysine / metabolism*
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Mice
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Mice, Knockout
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Protein Processing, Post-Translational*
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Sirtuin 1 / physiology*
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
Substances
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Sirt1 protein, mouse
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Sirtuin 1
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Lysine