PXD072723-1

PXD072723 is an original dataset announced via ProteomeXchange.

Title	One-Step Chemoenzymatic Labeling and Oxime-Reversible Enrichment for O-GlcNAc Profiling in Oxidative Stress
Description	Chemoenzymatic labeling has greatly advanced O-GlcNAc proteomics, yet existing strategies using azide-modified donors (e.g., UDP-GalNAz) require multi-step click-based workflows that introduce substantial sample loss and limit sensitivity toward low-abundance targets. While recent one-step labeling with biotinylated UDP-GalNAc donors streamlines workflows, the irreversible avidin–biotin interaction prevents glycopeptide recovery and restricts site-specific analysis. To address these challenges, we developed a ketone-reversible O-GlcNAc enrichment (KRO-GlcNAc) strategy employing a ketone-functionalized UDP-GalNAc analog (UDP-GalNLev) for one-step enzymatic tagging. GalT1(Y289L) transfers the GalNLev moiety with >96% efficiency, eliminating intermediate reactions and minimizing handling-induced loss. Crucially, reversible oxime-based capture on hydroxylamine-functionalized beads followed by mild methoxyamine release not only overcomes the irreversible nature of reported ketone-hydroxylamine-biotin enrichment, but fundamentally address the broader limitations that originally necessitated the transition from ketone-based labeling to azide chemistry. Application of KRO-GlcNAc to HeLa cells identified 4,129 potential O-GlcNAcylation sites across 740 proteins, with ~30% being novel. Functional studies revealed that O-GlcNAcylation regulates nuclear-cytoplasmic shuttling of chromatin remodeler EP400, linking this modification to stress granule assembly. This platform thus provides a powerful and versatile tool for advancing O-GlcNAcylation research.
HostingRepository	iProX
AnnounceDate	2026-01-06
AnnouncementXML	Submission_2026-01-06_16:37:25.739.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Li Liu
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF; Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2026-01-06 16:37:08	ID requested
⏵ 1	2026-01-06 16:37:26	announced

Dataset with its publication pending

submitter keyword: One-step Chemoenzymatic Labeling, O-GlcNAcylation, Glycopeptide Reversible Enrichment, Oxime Dynamic Formation, Oxidative Stress

Mingliang Ye
contact affiliation	Dalian Institute of Chemical Physics, Chinese Academy of Sciences
contact email	mingliang@dicp.ac.cn
lab head
Li Liu
contact affiliation	Dalian Institute of Chemical Physics, Chinese Academy of Sciences
contact email	leigh1809@dicp.ac.cn
dataset submitter

iProX dataset URI