biostudies-literatureUribe IRNational Institute of Allergy and Infectious DiseasesNICHD NIH HHSNIH Office of the DirectorNIAID NIH HHSNational Institute of General Medical SciencesNIGMS NIH HHSNational Institute of Child Health and Human DevelopmentNIH HHS1791-1803https://www.ebi.ac.uk/biostudies/studies/S-EPMC12282491biostudies-literatureUnknown24(4)ADP-ribosylation, characterized by the addition of adenosine diphosphate ribose, can occur in both monomeric (MARylation) and polymeric (PARylation) forms. Little is known about the specific contributions of MARylation and PARylation to cellular processes due to a lack of tools for jointly investigating these individual forms. We present a novel mass spectrometry (MS)-based proteomics approach that preserves information about the native ADP-ribosylation form associated with the modification site within a single proteomics experiment. Our workflow enables the simplified, binary identification of ADP-ribosylation forms, avoiding some challenges typically presented by PARylated peptides during MS analysis. Our method uses the coronaviral enzyme NS2 to reverse our previous labeling approach, EJournal of proteome researchdELTA-MS: A Mass Spectrometry-Based Proteomics Approach for Identifying ADP-Ribosylation Sites and Forms.PMC12282491R01 HD106051R01 GM104135R35GM147140R35 GM147140R01AI118891R01 AI118891R44 GM153000R01HD106051F31GM143918F31 GM143918S10 OD021844R01GM104135S10OD021844R44GM153000Dasovich MNa CHFilippov DVHunt SRLeung AKLZahn EGarcia BAOrsburn BCUribe IROnuoha UCSearfoss RVoorneveld JValadez CKim HBfalsedELTA-MS: A Mass Spectrometry-Based Proteomics Approach for Identifying ADP-Ribosylation Sites and Forms.ADP-ribosylation, characterized by the addition of adenosine diphosphate ribose, can occur in both monomeric (MARylation) and polymeric (PARylation) forms. Little is known about the specific contributions of MARylation and PARylation to cellular processes due to a lack of tools for jointly investigating these individual forms. We present a novel mass spectrometry (MS)-based proteomics approach that preserves information about the native ADP-ribosylation form associated with the modification site within a single proteomics experiment. Our workflow enables the simplified, binary identification of ADP-ribosylation forms, avoiding some challenges typically presented by PARylated peptides during MS analysis. Our method uses the coronaviral enzyme NS2 to reverse our previous labeling approach, E2025-01-01T00:00:00Z2025 Apr2026-03-27T15:14:37.627Z2025-08-23T03:09:03.569ZS-EPMC122824914007941510.1021/acs.jproteome.4c00890