Proteome-Wide Identification of In Vivo ADP-Ribose Acceptor Sites by Liquid Chromatography-Tandem Mass Spectrometry
Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › fagfællebedømt
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.
Originalsprog | Engelsk |
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Titel | Poly(ADP-Ribose) Polymerase |
Redaktører | Alexei V. Tulin |
Antal sider | 14 |
Vol/bind | 1608 |
Forlag | Humana Press |
Publikationsdato | 2017 |
Sider | 149-162 |
Kapitel | 11 |
ISBN (Trykt) | 978-1-4939-6992-0 |
ISBN (Elektronisk) | 978-1-4939-6993-7 |
DOI | |
Status | Udgivet - 2017 |
Navn | Methods in molecular biology (Clifton, N.J.) |
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ISSN | 1064-3745 |
ID: 187623120