biostudies-literatureMadsen TDDanmarks GrundforskningsfondNNF Center for Basic Metabolic ResearchLundbeckfondenNovo Nordisk FondenLundbeck Foundation4033https://www.ebi.ac.uk/biostudies/studies/S-EPMC7441158biostudies-literatureUnknown11(1)Peptide hormones and neuropeptides encompass a large class of bioactive peptides that regulate physiological processes like anxiety, blood glucose, appetite, inflammation and blood pressure. Here, we execute a focused discovery strategy to provide an extensive map of O-glycans on peptide hormones. We find that almost one third of the 279 classified peptide hormones carry O-glycans. Many of the identified O-glycosites are conserved and are predicted to serve roles in proprotein processing, receptor interaction, biodistribution and biostability. We demonstrate that O-glycans positioned within the receptor binding motifs of members of the neuropeptide Y and glucagon families modulate receptor activation properties and substantially extend peptide half-lives. Our study highlights the importancNature communicationsAn atlas of O-linked glycosylation on peptide hormones reveals diverse biological roles.PMC7441158R249-2017-1095NNF17OC0026030NNF12OC1016060Holst GroupDNRF107Schjoldager KTJoshi HJHolst JJHansen LHMartin CGoetze JPHintze JYe ZMadsen TDAndersen DBRosenkilde MMJebari SVakhrushev SYGoth CKJu TKuhre REfalseAn atlas of O-linked glycosylation on peptide hormones reveals diverse biological roles.Peptide hormones and neuropeptides encompass a large class of bioactive peptides that regulate physiological processes like anxiety, blood glucose, appetite, inflammation and blood pressure. Here, we execute a focused discovery strategy to provide an extensive map of O-glycans on peptide hormones. We find that almost one third of the 279 classified peptide hormones carry O-glycans. Many of the identified O-glycosites are conserved and are predicted to serve roles in proprotein processing, receptor interaction, biodistribution and biostability. We demonstrate that O-glycans positioned within the receptor binding motifs of members of the neuropeptide Y and glucagon families modulate receptor activation properties and substantially extend peptide half-lives. Our study highlights the importanc2020-01-01T00:00:00Z2020 Aug2026-04-16T23:16:03.427Z2020-09-05T07:10:35ZS-EPMC74411583282016710.1038/s41467-020-17473-1