Mapping Physiological ADP-Ribosylation Using Activated Ion Electron Transfer Dissociation

Sara C. Buch-Larsen, Ivo A. Hendriks, Jean M. Lodge, Martin Rykaer, Benjamin Furtwangler, Evgenia Shishkova, Michael S. Westphall, Joshua J. Coon, Michael L. Nielsen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

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 profile 9,000 ADPr peptides mapping to >5,000 unique ADPr sites from a limited number of cells exposed to oxidative stress and identify 120% and 28% more ADPr peptides compared to contemporary strategies using ETD and electron-transfer higher-energy collisional dissociation (EThcD), respectively. Under physiological conditions, AI-ETD identifies 450 ADPr sites on low-abundant proteins, including in vivo cysteine modifications on poly(ADP-ribosyl)polymerase (PARP) 8 and tyrosine modifications on PARP14, hinting at specialist enzymatic functions for these enzymes. Collectively, our data provide insights into the physiological regulation of ADPr.

Original languageEnglish
Article number108176
JournalCell Reports
Volume32
Issue number12
Number of pages17
ISSN2211-1247
DOIs
Publication statusPublished - 2020

Keywords

  • POLY(ADP-RIBOSYL)ATION
  • PARP-1
  • SITE
  • IDENTIFICATION
  • PROTEIN
  • ASSOCIATION
  • GENERATION
  • MECHANISM
  • ARGININE
  • PRODUCT

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