RT Journal Article
SR Electronic
T1 Mapping physiological ADP-ribosylation using Activated Ion Electron Transfer Dissociation (AI-ETD)
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.01.27.921650
DO 10.1101/2020.01.27.921650
A1 Sara C. Buch-Larsen
A1 Ivo A. Hendriks
A1 Jean M. Lodge
A1 Martin Rykær
A1 Benjamin Furtwängler
A1 Evgenia Shishkova
A1 Michael S. Westphall
A1 Joshua J. Coon
A1 Michael L. Nielsen
YR 2020
UL http://biorxiv.org/content/early/2020/01/28/2020.01.27.921650.abstract
AB ADP-ribosylation (ADPr) is a post-translational modification that plays pivotal roles in a wide range of cellular processes. Mass spectrometry (MS)-based analysis of ADPr under physiological conditions, without relying on genetic or chemical perturbation, has been hindered by technical limitations. Here, we describe the applicability of Activated Ion Electron Transfer Dissociation (AI-ETD) for MS-based proteomics analysis of physiological ADPr using our unbiased Af1521 enrichment strategy. To benchmark AI-ETD, we profiled 9,000 ADPr peptides mapping to >5,000 unique ADPr sites from a limited number of cells exposed to oxidative stress, corresponding to 120% and 28% more ADPr peptides compared to contemporary strategies using ETD and EThcD, respectively. Under physiological conditions AI-ETD identified 450 ADPr sites on low-abundant proteins, including in vivo cysteine auto-modifications on PARP8 and tyrosine auto-modifications on PARP14, hinting at specialist enzymatic functions for these enzymes. Collectively, our data provides new insights into the physiological regulation of ADP-ribosylation.