Proteome-Wide Identification of In Vivo ADP-Ribose Acceptor Sites by Liquid Chromatography-Tandem Mass Spectrometry

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ADP-ribosylation is a posttranslational modification (PTM) that affects a variety of cellular processes. In recent years, mass spectrometry (MS)-based proteomics has become a valuable tool for studying ADP-ribosylation. However, studying this PTM in vivo in an unbiased and sensitive manner has remained a difficult challenge. Here, we describe a detailed protocol for unbiased analysis of ADP-ribosylated proteins and their ADP-ribose acceptor sites under physiological conditions. The method relies on the enrichment of mono-ADP-ribosylated peptides using the macrodomain Af1521 in combination with liquid chromatography-high-resolution tandem MS (LC-MS/MS). The 5-day protocol explains the step-by-step enrichment and identification of ADP-ribosylated peptides from cell culture stage all the way through to data processing using the MaxQuant software suite.

Original languageEnglish
Title of host publicationPoly(ADP-Ribose) Polymerase
EditorsAlexei V. Tulin
Number of pages14
PublisherHumana Press
Publication date2017
ISBN (Print)978-1-4939-6992-0
ISBN (Electronic)978-1-4939-6993-7
Publication statusPublished - 2017
SeriesMethods in molecular biology (Clifton, N.J.)

    Research areas

  • Journal Article

ID: 187623120