biostudies-literatureGagne JPCanadian Institutes of Health Research7788-805https://www.ebi.ac.uk/biostudies/studies/S-EPMC3439892biostudies-literatureUnknown40(16)Upon DNA damage induction, DNA-dependent poly(ADP-ribose) polymerases (PARPs) synthesize an anionic poly(ADP-ribose) (pADPr) scaffold to which several proteins bind with the subsequent formation of pADPr-associated multiprotein complexes. We have used a combination of affinity-purification methods and proteomics approaches to isolate these complexes and assess protein dynamics with respect to pADPr metabolism. As a first approach, we developed a substrate trapping strategy by which we demonstrate that a catalytically inactive Poly(ADP-ribose) glycohydrolase (PARG) mutant can act as a physiologically selective bait for the isolation of specific pADPr-binding proteins through its macrodomain-like domain. In addition to antibody-mediated affinity-purification methods, we used a pADPr macrodomNucleic acids researchQuantitative proteomics profiling of the poly(ADP-ribose)-related response to genotoxic stress.PMC3439892MOP-74648MOP-14052Krietsch JBoutin MEthier CPoirier GGPic EPaquet EIsabelle MKelly IMoon KMFoster LJGagne JPfalseQuantitative proteomics profiling of the poly(ADP-ribose)-related response to genotoxic stress.Upon DNA damage induction, DNA-dependent poly(ADP-ribose) polymerases (PARPs) synthesize an anionic poly(ADP-ribose) (pADPr) scaffold to which several proteins bind with the subsequent formation of pADPr-associated multiprotein complexes. We have used a combination of affinity-purification methods and proteomics approaches to isolate these complexes and assess protein dynamics with respect to pADPr metabolism. As a first approach, we developed a substrate trapping strategy by which we demonstrate that a catalytically inactive Poly(ADP-ribose) glycohydrolase (PARG) mutant can act as a physiologically selective bait for the isolation of specific pADPr-binding proteins through its macrodomain-like domain. In addition to antibody-mediated affinity-purification methods, we used a pADPr macrodom2012-01-01T00:00:00Z2012 Sep2026-04-29T13:32:56.652Z2019-03-27T00:57:51ZS-EPMC34398922266991110.1093/nar/gks486