Dataset Information

HeLa, HEK293T and mouse lung, heart and spleen LC-MS/MS

ABSTRACT: Proteins are ubiquitously modified with glycans of varied chemical structures via distinct glycosidic linkages, making the landscape of protein glycosylation challenging to map. Profiling of intact glycopeptides with mass spectrometry (MS) has recently emerged as a powerful tool for revealing matched information of the glycosylation sites and attached glycans (i.e., intact glycosites), which, however, is largely limited to individual glycosylation types. Here, we develop Click-iG, which integrates metabolic labeling of glycans with clickable unnatural sugars, an optimized MS method, and a tailored version of pGlyco3 software to enable simultaneous enrichment and profiling of three types of—N-linked, O-linked, and O-GlcNAcylated—intact glycopeptides. We demonstrated Click-iG by identification of thousands of intact glycosites in cell lines and living mice. From the mouse lung, heart, and spleen, a total of 547 N-glycosylation sites with 161 glycan compositions, 198 O-glycosylation sites with 262 glycans, and 1,947 O‑GlcNAcylation sites were identified. The Click-iG-enabled unprecedent coverage on the protein glycosylation landscape lays the foundation for interrogating crosstalk between different glycosylation pathways.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Heart

SUBMITTER: Jialin Liu

LAB HEAD: xing chen

PROVIDER: PXD039144 | Pride | 2025-02-12

REPOSITORIES: pride

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Dataset's files

Source:

			Action	DRS
	DBDB-N.txt	Txt
	DBDB-O.txt	Txt
	Lumos3_HomeMadeCol_ChenXing_LJL_WT-heart_75um_50cm_360min_HCD30_stepped10_PDEThcD35_20221212_F1_R1.raw	Raw
	Lumos3_HomeMadeCol_ChenXing_LJL_WT-heart_75um_50cm_360min_HCD30_stepped10_PDEThcD35_20221212_F1_R2.raw	Raw
	Lumos3_HomeMadeCol_ChenXing_LJL_WT-heart_75um_50cm_360min_HCD30_stepped10_PDEThcD35_20221212_F1_R3.raw	Raw

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Publications

Click-iG: Simultaneous Enrichment and Profiling of Intact N-linked, O-GalNAc, and O-GlcNAcylated Glycopeptides.

Liu Jialin J Cheng Bo B Fan Xinqi X Zhou Xinyue X Wang Jiankun J Zhou Wen W Li Hengyu H Zeng Wenfeng W Yang Pengyuan P Chen Xing X

Angewandte Chemie (International ed. in English) 20230725 36

Proteins are ubiquitously modified with glycans of varied chemical structures through distinct glycosidic linkages, making the landscape of protein glycosylation challenging to map. Profiling of intact glycopeptides with mass spectrometry (MS) has recently emerged as a powerful tool for revealing matched information of the glycosylation sites and attached glycans (i.e., intact glycosites), but is largely limited to individual glycosylation types. Herein, we describe Click-iG, which integrates me ...[more]

PMID: 37431278

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