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

ABSTRACT: Blood and plasma proteins are heavily investigated as biomarkers for different diseases. However, the post-translational modifications of these proteins are rarely analyzed since blood contains many enzymes that rapidly remove the 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 established that plasma phosphodiesterases and ADP-ribosylhydrolases rapidly demodified 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.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Sus Scrofa Domesticus (domestic Pig)

TISSUE(S): Blood Plasma, Epithelial Cell, Blood

SUBMITTER: Deena Leslie Pedrioli  

LAB HEAD: Michael O. Hottiger, Prof. PhD, DVM

PROVIDER: PXD022156 | Pride | 2021-05-26

REPOSITORIES: Pride