Proteome-Wide Identification of In Vivo ADP-Ribose Acceptor Sites by Liquid Chromatography-Tandem Mass Spectrometry
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Proteome-Wide Identification of In Vivo ADP-Ribose Acceptor Sites by Liquid Chromatography-Tandem Mass Spectrometry. / Larsen, Sara C; Leutert, Mario; Bilan, Vera; Martello, Rita; Jungmichel, Stephanie; Young, Clifford; Hottiger, Michael O; Nielsen, Michael L.
Poly(ADP-Ribose) Polymerase. ed. / Alexei V. Tulin. Vol. 1608 Humana Press, 2017. p. 149-162 (Methods in molecular biology (Clifton, N.J.)).Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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TY - CHAP
T1 - Proteome-Wide Identification of In Vivo ADP-Ribose Acceptor Sites by Liquid Chromatography-Tandem Mass Spectrometry
AU - Larsen, Sara C
AU - Leutert, Mario
AU - Bilan, Vera
AU - Martello, Rita
AU - Jungmichel, Stephanie
AU - Young, Clifford
AU - Hottiger, Michael O
AU - Nielsen, Michael L
PY - 2017
Y1 - 2017
N2 - 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.
AB - 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.
KW - Journal Article
U2 - 10.1007/978-1-4939-6993-7_11
DO - 10.1007/978-1-4939-6993-7_11
M3 - Book chapter
C2 - 28695509
SN - 978-1-4939-6992-0
VL - 1608
T3 - Methods in molecular biology (Clifton, N.J.)
SP - 149
EP - 162
BT - Poly(ADP-Ribose) Polymerase
A2 - Tulin, Alexei V.
PB - Humana Press
ER -
ID: 187623120