| Literature DB >> 28695509 |
Sara C Larsen1, Mario Leutert2,3, Vera Bilan2,3, Rita Martello1, Stephanie Jungmichel1, Clifford Young1, Michael O Hottiger2, Michael L Nielsen4.
Abstract
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.Entities:
Keywords: ADP-ribosylation; ADP-ribosylome; Af1521 macrodomain enrichment; Affinity purification; Mass spectrometry; PARG; Proteomics
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Year: 2017 PMID： 28695509 DOI： 10.1007/978-1-4939-6993-7_11
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745