%0 Journal Article %A Stephanie C. Lüthi %A Anna Howald %A Kathrin Nowak %A Robert Graage %A Giody Bartolomei %A Christine Neupert %A Xaver Sidler %A Deena M. Leslie Pedrioli %A Michael O. Hottiger %T Development of a mass-spectrometry based method for the identification of the in vivo whole blood and plasma ADP-ribosylomes %D 2020 %R 10.1101/2020.11.17.384719 %J bioRxiv %P 2020.11.17.384719 %X Blood and plasma proteins are heavily investigated as biomarkers for different diseases. However, the post-translational modification states of these proteins are rarely analyzed since blood contains many enzymes that rapidly remove these modification after sampling. In contrast to the well-described role of protein ADP-ribosylation in cells and organs, its role in blood remains mostly uncharacterized. Here, we discovered that plasma phosphodiesterases and/or ADP-ribosylhydrolases rapidly demodify in vitro ADP-ribosylated proteins. Thus, to identify the in vivo whole blood and plasma ADP-ribosylomes, we established a novel mass-spectrometry based workflow that was applied to blood samples collected from LPS-treated pigs (Sus scrofa), which serves as a model for human systemic inflammatory response syndrome. These analyses identified 60 ADP-ribosylated proteins, 17 of which were ADP-ribosylated plasma proteins. This new protocol provides an important step forward for the rapidly developing field of ADP-ribosylation and defines the blood and plasma ADP-ribosylomes under both healthy and disease conditions.Competing Interest StatementThe authors have declared no competing interest. %U https://www.biorxiv.org/content/biorxiv/early/2020/11/17/2020.11.17.384719.full.pdf