New Quantitative Mass Spectrometry Approaches Reveal Different ADP-ribosylation Phases Dependent On the Levels of Oxidative Stress

作者：Vera Bilan, Nathalie Selevsek, Hans A. V. Kistemaker, Jeannette Abplanalp, Roxane Feurer, Dmitri V. Filippov, Michael O. Hottiger · 发表于：Molecular & Cellular Proteomics · 年份：2017 · DOI：10.1074/mcp.o116.065623 · 被引用次数：46 · 研究领域：PARP inhibition in cancer therapy、DNA Repair Mechanisms、Mass Spectrometry Techniques and Applications

Oxidative stress is a potent inducer of protein ADP-ribosylation. Although individual oxidative stress-induced ADP-ribosylated proteins have been identified, it is so far not clear to which extent different degrees of stress severity quantitatively and qualitatively alter ADP-ribosylation. Here, we investigated both quantitative and qualitative changes of the hydrogen peroxide (H 2 O 2 )-induced ADP-ribosylome using a label-free shotgun quantification and a parallel reaction monitoring (PRM) mass spectrometry approach for a selected number of identified ADP-ribosylated peptides. Although the major part of the basal HeLa ADP-ribosylome remained unchanged upon all tested H 2 O 2 concentrations, some selected peptides change the extent of ADP-ribosylation depending on the degree of the applied oxidative stress. Low oxidative stress ( i.e. 4 μm and 16 μm H 2 O 2 ) caused a reduction in ADP-ribosylation of modified proteins detected under untreated conditions. In contrast, mid to strong oxidative stress (62 μm to 1 mm H 2 O 2 ) induced a significant increase in ADP-ribosylation of oxidative stress-targeted proteins. The application of the PRM approach to SKOV3 and A2780, ovarian cancer cells displaying different sensitivities to PARP inhibitors, revealed that the basal and the H 2 O 2 -induced ADP-ribosylomes of SKOV3 and A2780 differed significantly and that the sensitivity to PARP inhibitors correlated with the level of ARTD1 expression in these cells. Overall, this new PRM-MS a...