Journal Article

Proteome-wide identification of the endogenous ADP-ribosylome of mammalian cells and tissue

Jungmichel, Stephanie, Bilan, Vera, Larsen, Sara C., Martello, Rita, Young, Clifford, Hottiger, Michael O., Leutert, Mario, Nielsen, Michael L.

2016

Overview

Although protein ADP-ribosylation is involved in diverse biological processes, it has remained a challenge to identify ADP-ribose acceptor sites. Here, we present an experimental workflow for sensitive and unbiased analysis of endogenous ADP-ribosylation sites, capable of detecting more than 900 modification sites in mammalian cells and mouse liver. In cells, we demonstrate that Lys residues, besides Glu, Asp and Arg residues, are the dominant in vivo targets of ADP-ribosylation during oxidative stress. In normal liver tissue, we find Arg residues to be the predominant modification site. The cellular distribution and biological processes that involve ADP-ribosylated proteins are different in cultured cells and liver tissue, in the latter of which the majority of sites were found to be in cytosolic and mitochondrial protein networks primarily associated with metabolism. Collectively, we describe a robust methodology for the assessment of the role of ADP-ribosylation and ADP-ribosyltransferases in physiological and pathological states. ADP-ribosylation is a reversible post-translational protein modification involved in many cellular processes. Here the authors describe a sensitive approach for the analysis of ADP-ribosylation sites under physiologic conditions and identify lysine residues as in vivo targets of ADP-ribosylation.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 631/1647/296 / 631/337/458/2389 / Adenosine diphosphate / Amino acids / Humanities and Social Sciences / Kinases / Life sciences / Liver / Mammals / multidisciplinary / Oxidative stress / Peptides / Physiology / Proteins / Science / Science (multidisciplinary)