Common response
Under the inhibition of protein N-glycosylation, O-GlcNAcylated proteins related to stress response and translation are commonly changed in different types of cells.
Analytical Chemistry 2026, 98, 15689–15699
Systematic Quantification of Protein O-GlcNAcylation Reveals Common and Cell-Type-Specific Responses to N-Glycosylation Inhibition in Human Cells
Both protein O-GlcNAcylation and N-glycosylation are extremely important in human cells and regulate many cellular events. While O-GlcNAcylation is known to act as a stress sensor, its changes in human cells with N-glycosylation perturbations remain to be explored. In this study, we comprehensively and site-specifically studied common and cell-type-specific responses of protein O-GlcNAcylation under N-glycosylation inhibition in three types of human cells (HEK293T, HepG2, and Jurkat cells) by integrating metabolic labeling, bio-orthogonal chemistry, and multiplexed proteomics.
Each point is an O-GlcNAcylated protein. Red = up under Tm, blue = down (|log₂FC| > 0.5, adj. P < 0.05). Click any panel to open the full browser.
Cell-type specificity
O-GlcNAcylated proteins related to leukocyte proliferation and T-cell activation were upregulated in Jurkat cells, while in HEK293T cells, those associated with ribonucleotide metabolism and ribosome biogenesis were upregulated.
Local structure
Site-specific analysis revealed that O-GlcNAcylation sites in structured regions exhibited larger abundance changes compared with those in intrinsically disordered regions.
O-GlcNAc proteins
Tunicamycin vs. control, per cell type. Click a row for its TMT profile.
From the paperUnder N-glycosylation inhibition, O-GlcNAcylated proteins related to stress response and translation are commonly changed across the three cell types, while the responses of protein O-GlcNAcylation are also highly specific to individual cell types.
O-GlcNAc sites
Site-level changes with structural context. Select a site to view it on its AlphaFold model.
From the paperSite-specific analysis revealed that O-GlcNAcylation sites in structured regions exhibited larger abundance changes compared with those in intrinsically disordered regions.
log₂FC by local structure — structured-region sites (helix/sheet) vs. intrinsically disordered.
Select a site to load its AlphaFold structure.
O-GalNAc
Mucin-type O-GalNAc proteins and sites captured alongside O-GlcNAc.
From the paperA total of 485 O-GalNAcylated (Tn antigen) proteins were captured across the three cell types. Under N-glycosylation inhibition, the abundance of O-GalNAcylated proteins in the classical secretory pathway decreased, and these downregulated proteins were involved in ER proteostasis, glycoprotein maturation, and receptor regulation.
Whole proteome
Total protein-abundance change — the background against which glyco changes are read. Loaded on demand.
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Annotated spectra
Representative HCD / EThcD glycopeptide spectra behind the localized sites. The full set is on PRIDE.
Data & downloads
Everything here is derived from the deposited data and the published Supporting Information.