PXD029482-1
PXD029482 is an original dataset announced via ProteomeXchange.
Dataset Summary
Title | Comprehensive O-GlcNAc glycoproteomics on NOTCH1 EGF repeats refined sequons for O-GlcNAcylation and uncovered unique Lewis X epitopes in mammals |
Description | The O-GlcNAc modification of Notch receptors regulates Notch ligand interactions in a manner distinct from other forms of O-glycans on epidermal growth factor-like (EGF) repeats of Notch receptors. Although many proteins, besides Notch receptors, are expected to be O-GlcNAcylated by EGF domain-specific O-GlcNAc transferase (EOGT), only a small number of proteins have been reported to be modified in vivo, and elongated O-GlcNAc glycans have not been extensively explored. To extend our view of the specificity and variety of the glycan modification, we conducted a comprehensive analysis of O-GlcNAc glycans on NOTCH1 in mammals. Mass spectrometric analysis of NOTCH1 fragments expressed in HEK293T cells revealed that several EGF domains with putative O-GlcNAcylation sites were hardly modified with O-GlcNAc. Although amino acid residues before the modification site are preferentially occupied with aromatic residues, Phe and Tyr are preferrable to Trp for the apparent modification with O-GlcNAc. Furthermore, a minor form of fucosylated O-GlcNAc glycans was detected in a subset of EGF domains. Fucosylation of O-GlcNAc glycans was enhanced by FUT1, FUT2, or FUT9 expression. The FUT9-dependent Lewis X epitope was confirmed by immunoblotting using an anti-Lewis X antibody. As expected from the similarity in the glycan structures, the Lexis X antigen was detected on O-fucose glycans. Notably, the Lewis X structure on O-glycans was identified in endogenous NOTCH1 isolated from MCF7 cells. Our results refined the putative consensus sequence for the EOGT-dependent extracellular O-GlcNAc modification in mammals and revealed the structural diversity of functional Notch O-glycans. |
HostingRepository | jPOST |
AnnounceDate | 2022-11-02 |
AnnouncementXML | Submission_2022-11-01_08:00:05.216.xml |
DigitalObjectIdentifier | |
ReviewLevel | Non peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Yohei Tsukamoto |
SpeciesList | scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; |
ModificationList | S-carboxamidomethyl-L-cysteine; L-methionine sulfoxide; unknown modification |
Instrument | LTQ Orbitrap |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
---|---|---|---|
0 | 2021-11-01 09:21:34 | ID requested | |
⏵ 1 | 2022-11-01 08:00:05 | announced |
Publication List
Dataset with its publication pending |
Keyword List
submitter keyword: O-GlcNAc |
Contact List
Tetsuya Okajima | |
---|---|
lab head | |
Yohei Tsukamoto | |
contact affiliation | Nagoya University |
dataset submitter |
Full Dataset Link List
jPOST dataset URI |
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