Development of a mass-spectrometry based method for the identification of the in vivo whole blood and plasma ADP-ribosylomes

作者：Stephanie C. Lüthi, Anna Howald, Kathrin Nowak, Robert Graage, Giody Bartolomei, Christine Neupert, Xaver Sidler, Deena M. Leslie Pedrioli, Michael O. Hottiger · 发表于：bioRxiv (Cold Spring Harbor Laboratory) · 年份：2020 · DOI：10.1101/2020.11.17.384719 · 研究领域：PARP inhibition in cancer therapy、Cardiac electrophysiology and arrhythmias、Calcium signaling and nucleotide metabolism

ABSTRACT 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.