biostudies-literaturePiovesana SEuropean Research CouncilSapienza Universit? di Roma9234-9241https://www.ebi.ac.uk/biostudies/studies/S-EPMC9260711biostudies-literatureUnknown94(26)Site localization of protein sulfation by high-throughput proteomics remains challenging despite the technological improvements. In this study, sequence analysis and site localization of sulfation in tryptic peptides were determined under a conventional nano-liquid chromatography-mass spectrometry configuration. Tryptic sulfopeptide standards were used to study different fragmentation strategies, including collision-induced dissociation (CID), higher-energy collisional dissociation (HCD), electron-transfer dissociation (ETD), electron-transfer/higher-energy collision dissociation (EThcD), and electron-transfer/collision-induced dissociation (ETciD), in the positive ionization mode. Sulfopeptides displayed only neutral loss of SO₃ under CID, while the sequence could be determinedAnalytical chemistryThe Key Role of Metal Adducts in the Differentiation of Phosphopeptide from Sulfopeptide Sequences by High-Resolution Mass Spectrometry.PMC9260711AR220172B85F20D6823839Cerrato ALagana AMontone CMPiovesana SZenezini Chiozzi RCavaliere CCapriotti ALfalseThe Key Role of Metal Adducts in the Differentiation of Phosphopeptide from Sulfopeptide Sequences by High-Resolution Mass Spectrometry.Site localization of protein sulfation by high-throughput proteomics remains challenging despite the technological improvements. In this study, sequence analysis and site localization of sulfation in tryptic peptides were determined under a conventional nano-liquid chromatography-mass spectrometry configuration. Tryptic sulfopeptide standards were used to study different fragmentation strategies, including collision-induced dissociation (CID), higher-energy collisional dissociation (HCD), electron-transfer dissociation (ETD), electron-transfer/higher-energy collision dissociation (EThcD), and electron-transfer/collision-induced dissociation (ETciD), in the positive ionization mode. Sulfopeptides displayed only neutral loss of SO₃ under CID, while the sequence could be determined2022-01-01T00:00:00Z2022 Jul2025-04-05T14:25:58.229Z2022-07-23T09:39:22.488ZS-EPMC92607113571406210.1021/acs.analchem.1c05621