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Large-scale and Site-specific Mapping of Intact O-glycans on Glycopeptides with IMPa

ABSTRACT: We employed the newly available IMPa O-glycoprotease from Pseudomonas aeruginosa for O-glycoproteomics analysis of cultured cells and tissues. The glycopeptides were extracted, purified, and conjugated to a solid support before an enzymatic cleavage by IMPa. O-glycopeptides were analyzed by EThcD, which allowed for a site-specific and global analysis of O-glycans, especially those sialylated and the Tn antigen. Through wild-type or SimpleCell HEK293 cells, we present two approaches, one for the analysis of total O-glycoproteome and the other for the Tn glycoproteome, and showed that IMPa and EThcD are necessary for confident localization of O-glycans on glycopeptides. We applied this method to study the O-glycoproteome of mouse brain, which revealed the high frequency of sialylated O-glycans and the Tn antigen on brain glycoproteins.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Brain, Kidney

SUBMITTER: Suttipong Suttapitugsakul

LAB HEAD: Richard D. Cummings

PROVIDER: PXD037415 | Pride | 2023-08-28

REPOSITORIES: Pride

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			Action	DRS
	SS_03272023_mouse_brain_N_r1.raw	Raw
	SS_03272023_mouse_brain_N_r1.txt	Txt
	SS_03272023_mouse_brain_N_r2.raw	Raw
	SS_03272023_mouse_brain_N_r2.txt	Txt
	SS_03272023_mouse_brain_N_r3.raw	Raw

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