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PXD031297

PXD031297 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleO-Fucose glycans on mouse NOTCH1 from activated T cells of Fringe mutant mice
DescriptionNOTCH1 (N1) is a transmembrane receptor that initiates a cell-cell signaling pathway controlling various cell fate specifications in metazoans. The addition of O-fucose by Protein O-fucosyltransferase 1 (POFUT1) to Epidermal Growth Factor-like (EGF) repeats in the N1 extracellular domain is essential for N1 function, and modification of O-fucose with GlcNAc by the Fringe family of glycosyltransferases modulates Notch activity. Prior cell-based studies showed that POFUT1 modifies EGF repeats containing the appropriate consensus sequence at high stoichiometry, while Fringe GlcNAc-transferases (LFNG, MFNG and RFNG) modify O-fucose on only a subset of NOTCH1 EGF repeats. Previous in vivo studies showed that each FNG affects naïve T cell development. To examine Fringe modifications of N1 expressed at a physiological level, we used mass spectral glycoproteomic methods to analyze O-fucose glycans of endogenous N1 from activated T cells obtained from mice lacking all Fringe enzymes, or expressing only a single FNG. While most O-fucose sites were modified at high stoichiometry, only EGF6, EGF16, EGF26, and EGF27 were extended in control T cells. Cell-based assays of N1 lacking fucose at each of those O-fucose sites revealed minor functional correlations in the EGF16 and EGF27 mutant with Notch ligand binding. In activated T cells expressing only LFNG, MFNG or RFNG alone, the extension of O-fucose with GlcNAc in the same EGF repeats was diminished, consistent with cooperative interactions when all three Fringes were present. The combined data open the door for the analysis of O-glycans on endogenous N1 derived from different cell types.
HostingRepositoryPRIDE
AnnounceDate2022-08-12
AnnouncementXMLSubmission_2022-08-11_23:58:12.077.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterKenjiroo Matsumoto
SpeciesList scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationListmonohydroxylated residue; iodoacetic acid derivatized residue
InstrumentQ Exactive
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-01-27 01:44:24ID requested
12022-08-11 23:58:12announced
Publication List
Matsumoto K, Kumar V, Varshney S, Nairn AV, Ito A, Pennarubia F, Moremen KW, Stanley P, Haltiwanger RS, Fringe GlcNAc-transferases differentially extend O-fucose on endogenous NOTCH1 in mouse activated T cells. J Biol Chem, 298(7):102064(2022) [pubmed]
Keyword List
submitter keyword: NOTCH1, O-fucose, POFUT1, Fringe, T cell
Contact List
Kenjiroo Matsumoto
contact affiliationComplex Carbohydrate Research Center
contact emailkmatsumoto@uga.edu
lab head
Kenjiroo Matsumoto
contact affiliationComplex Carbohydrate Research Center
contact emailkenjiroo.matsumoto@gmail.com
dataset submitter
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Dataset FTP location
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