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PXD020077

PXD020077 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleElectron-based dissociation is needed for O-glycopeptides derived from OpeRATOR proteolysis
DescriptionThe recently described O-glycoprotease OpeRATOR presents exciting opportunities for O-glycoproteomics. This bacterial enzyme purified from A. muciniphila cleaves N-terminally to serine and threonine residues that are modified with (preferably asialylated) O-glycans, providing orthogonal cleavage relative to canonical proteases (e.g., trypsin) to improve O-glycopeptide characterization with tandem mass spectrometry (MS/MS). O-glycopeptides with a modified N-terminal residue, such as those generated by OpeRATOR, present several potential benefits, perhaps the most notable being de facto O-glycosite localization without the need of glycan-retaining fragments in MS/MS spectra. Indeed, O-glycopeptides modified exclusively at the N-terminus would enable O-glycoproteomic methods to rely solely on collision-based fragmentation rather than electron-driven dissociation, but modified peptides would need to reliably contain only a single O-glycosite. Here we characterize the number of O-glycosites (i.e., missed cleavages) that are present in OpeRATOR-derived O-glycopeptides using methods that combine collision- and electron-based fragmentation. Our data show that over 50% of O-glycopeptides generated from combined digestion using OpeRATOR and trypsin contain multiple O-glycosites, indicating that collision-based fragmentation is not sufficient for OpeRATOR-centric studies and that electron-driven dissociation remains a requirement for site-specific O-glycopeptide characterization.
HostingRepositoryPRIDE
AnnounceDate2020-11-03
AnnouncementXMLSubmission_2020-11-02_22:58:23.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterNicholas Riley
SpeciesList scientific name: Bos taurus (Bovine); NCBI TaxID: 9913; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListcomplex glycosylation
InstrumentOrbitrap Fusion
Dataset History
RevisionDatetimeStatusChangeLog Entry
02020-06-29 00:11:24ID requested
12020-11-02 22:58:24announced
Publication List
Riley NM, Malaker SA, Bertozzi CR, Electron-Based Dissociation Is Needed for O-Glycopeptides Derived from OpeRATOR Proteolysis. Anal Chem, 92(22):14878-14884(2020) [pubmed]
Keyword List
submitter keyword: glycoproteomics, electron transfer dissociation, O-glycosites, OpeRATOR, protease
Contact List
Carolyn R. Bertozzi
contact affiliation1) Department of Chemistry, Stanford University, Stanford, California, USA 2) Howard Hughes Medical Institute, Stanford, California, USA
contact emailbertozzi@stanford.edu
lab head
Nicholas Riley
contact affiliationStanford University
contact emailriley.nicholasm@gmail.com
dataset submitter
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Dataset FTP location
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