Proteome-Wide Identification of In Vivo ADP-Ribose Acceptor Sites by Liquid Chromatography-Tandem Mass Spectrometry
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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 language | English |
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Title of host publication | Poly(ADP-Ribose) Polymerase |
Editors | Alexei V. Tulin |
Number of pages | 14 |
Volume | 1608 |
Publisher | Humana Press |
Publication date | 2017 |
Pages | 149-162 |
Chapter | 11 |
ISBN (Print) | 978-1-4939-6992-0 |
ISBN (Electronic) | 978-1-4939-6993-7 |
DOIs | |
Publication status | Published - 2017 |
Series | Methods in molecular biology (Clifton, N.J.) |
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ISSN | 1064-3745 |
- Journal Article
Research areas
ID: 187623120