Identification of ADP-ribosylation targets in living HeLa cells using cell-permeable NAD+ probes

ABSTRACT: Posttranslational modifications of proteins (PTMs) greatly enhance their functional diversity, surpassing the number of gene-encoded variations. One intriguing PTM is ADP-ribosylation, which utilizes nicotinamide adenine dinucleotide (NAD+) as a substrate and is essential in cell signaling pathways regulating cellular responses. Here, we report the first cell-permeable NAD+ analogs that are modified with a dye or an affinity tag, respectively, and demonstrate their suitability as tools for investigating of the cellular ADP-ribosylation process. Utilizing desthiobiotin-tagged probes, we characterized ADP-ribosylome changes during oxidative stress in HeLa cells. We identified proteins previously described as ADP-ribosylation targets or closely associated with them. Therefore, we believe that our cell-permeable NAD+ probes offer reliable tools for comprehensive ADP-ribosylation investigations and understanding cellular responses to stress.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Hela Cell

SUBMITTER: Renata Kasprzyk  

LAB HEAD: Andreas Marx

PROVIDER: PXD051967 | Pride | 2024-10-11

REPOSITORIES: Pride