MassIVE MSV000080334

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Oxidative stress induced ADP-ribosylome

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Description

Oxidative stress is a potent inducer of protein ADP-ribosylation. Although the proteins modified under oxidative stress have been identified, it is not clear, whether the number of modified proteins and/or the number ADP-ribosylation sites varies with stress intensity. Here, we investigated both the qualitative and quantitative changes in the HeLa cell ADP-ribosylome induced by mild (4 - 16 uM), moderate (64 - 250 uM), or severe, but non-lethal (1 mM) H2O2 concentrations using shotgun and parallel reaction monitoring mass spectrometry (PRM-MS). After moderate and severe oxidative stress, approximately 50% of proteins not ADP-ribosylated under basal conditions were induced, while ADP-ribosylation of the other 50% already ADP-ribosylated under basal conditions remained constant. In contrast, mild oxidative stress caused a reduction in detectable ADP-ribosylation of many proteins, which were induced under more severe oxidative stress. Overall, the number of ADP-ribosylation sites modified/de-modified did not change significantly under the various degrees of oxidative stress. Moreover, we applied the PRM method to study protein ADP-ribosylation in ovarian cancer cells that display different sensitivities to PARP inhibitors. These studies revealed similar ADP-ribosylation responses following H2O2 treatment, which, however, correlated in extent with ARTD1 abundance in these cells. Overall, our new MS approaches have proven to be highly useful in monitoring cellular protein ADP-ribosylation and have revealed unexpected fluctuations in proteins ADP-ribosylation depending on the degree of oxidative stress. [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Oxidative stress, ADP-ribosylation, Af1521 enrichment, HeLa, PRM, LFQ

Contact

Principal Investigators:
(in alphabetical order) 		Prof. Michael Hottiger
Submitting User: verabilan

Publications

Bilan V, Selevsek N, Kistemaker HAV, Abplanalp J, Feurer R, Filippov DV, Hottiger MO.
New Quantitative Mass Spectrometry Approaches Reveal Different ADP-ribosylation Phases Dependent On the Levels of Oxidative Stress.
Mol. Cell Proteomics. 2017 May;16(5):949-958. Epub 2017 Mar 21.

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