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

TY - JOUR

T1 - Proteome-wide identification of the endogenous ADP-ribosylome of mammalian cells and tissue

AU - Martello, Rita

AU - Leutert, Mario

AU - Jungmichel, Stephanie

AU - Bilan, Vera

AU - Larsen, Sara C

AU - Young, Clifford

AU - Hottiger, Michael O

AU - Nielsen, Michael L

PY - 2016/9/30

Y1 - 2016/9/30

N2 - 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.

AB - 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.

U2 - 10.1038/ncomms12917

DO - 10.1038/ncomms12917

M3 - Journal article

C2 - 27686526

SN - 2041-1723

VL - 7

JO - Nature Communications

JF - Nature Communications

M1 - 12917

ER -