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PXD004559

PXD004559 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleMapping and quantification of over 2,000 O-linked glycopeptides in activated human T cells with isotope-targeted glycoproteomics
DescriptionPost-translational modifications (PTMs) on proteins often function to regulate signaling cascades, with the activation of T cells during an adaptive immune response being a classic example. Mounting evidence indicates that the modification of proteins by O-linked Nacetylglucosamine (O-GlcNAc), the only mammalian glycan found on nuclear and cytoplasmic proteins, helps regulate T cell activation. Yet, a mechanistic understanding of how O-GlcNAc functions in T cell activation remains elusive, partly because of the difficulties in mapping and quantifying O-GlcNAc sites. Thus, to advance insight into the role of O-GlcNAc in T cell activation, we performed extensive glycosite mapping studies via direct glycopeptide measurement on resting and activated primary human T cells with a technique termed isotope targeted glycoproteomics. This approach led to the identification of over 2,000 intact O-GlcNAccontaining glycopeptides across 1,046 glycoproteins. A significant proportion (>45%) of the identified O-GlcNAc sites lie in close proximity to or coincide with known phosphorylation sites, supporting the potential for PTM crosstalk. Consistent with other studies, we find that O-GlcNAc sites in T cells lack a strict consensus sequence. To validate our results, we employed gel shift assays based on conjugating mass tags to O-GlcNAc groups. Notably, we observed that the transcription factors c-JUN and JUNB show higher levels of O-GlcNAc glycosylation and higher levels of expression in activated T cells. Overall, our findings provide a quantitative characterization of O-GlcNAc glycoproteins and their corresponding modification sites in primary human T cells, which will facilitate mechanistic studies into the function of O-GlcNAc in T cell activation.
HostingRepositoryPRIDE
AnnounceDate2018-01-25
AnnouncementXMLSubmission_2018-01-25_01:24:47.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterChristina Woo
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListN-acetylaminoglucosylated
InstrumentLTQ Orbitrap Elite
Dataset History
RevisionDatetimeStatusChangeLog Entry
02016-07-12 01:59:22ID requested
12018-01-25 01:24:48announced
Publication List
Woo CM, Lund PJ, Huang AC, Davis MM, Bertozzi CR, Pitteri SJ, Mapping and Quantification of Over 2000 O-linked Glycopeptides in Activated Human T Cells with Isotope-Targeted Glycoproteomics (Isotag). Mol Cell Proteomics, 17(4):764-775(2018) [pubmed]
Keyword List
submitter keyword: O-linked N-acetylglucosamine, O-GlcNAc, glycoproteomics, T cells, mass spectrometry
Contact List
Christina Woo
contact affiliationDepartment of Chemistry and Chemical Biology Harvard University
contact emailcwoo@chemistry.harvard.edu
lab head
Christina Woo
contact affiliationHarvard University
contact emailcwoo@chemistry.harvard.edu
dataset submitter
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