Profiling of the ADP?Ribosylome in Living Cells

Post-translational modification (PTM) with ADP-ribose and poly(ADP-ribose) using nicotinamide adenine dinucleotide (NAD+) as substrate is involved in the regulation of numerous cellular pathways eukaryotes, notably response to DNA damage caused by stress. Nevertheless, due intrinsic properties NAD+ e.g., high polarity associated poor cell passage, these PTMs are difficult characterize cells. Here, two new derivatives presented, which carry either a fluorophore or an affinity tag and, combination developed methods for mild delivery, allow studies living human We show that this approach allows not only imaging ADP-ribosylation cells but also proteome-wide analysis adaptation protein consequence environmental changes such H2O2-induced oxidative stress effect approved anti-cancer drug olaparib. Our results therefore pave way further functional clinical ADP-ribosylated proteome health disease.