Dataset Information

Modulation of Core-Fucosylation Oxidation for Selective Enrichment and Characterization of Cell Surface Glycoprotein

ABSTRACT: Protein core-fucosylation plays a crucial role in regulating cell surface protein functions and is involved in various biological processes, including cell signaling, immune response, and cancer progression. However, core-fucosylation (CF) poses particular challenges in identification and characterization due to its low abundance and the high heterogeneity of N-glycosylation. To overcome these challenges, this study systematically investigated the oxidation mechanisms of core-fucose and developed a selective enrichment strategy that combines chemical oxidation with glycan truncation for cell surface core-fucosylation characterization. Specifically, sodium periodate was employed to selectively oxidize cell surface glycans. In combination with the endoglycosidases Endo M and Endo F3, which possessed complementary substrate specificity and broad tolerance, this approach efficiently truncated N-glycans and leaving core-fucosylated glycopeptides bearing aldehyde tags. Utilizing reversible hydrazide chemistry, core-fucosylated glycopeptides were selectively enriched. The developed strategy was applied to profile cell surface core-fucosylatd protein in HeLa cells, which yielded 74 core-fucosylated glycopeptides corresponding to 21 key cell surface drug targets, thereby validating the efficacy of this approach. Collectively, this study systematically investigatd the mechnism of core-fucosylation oxidation and developed a new technical tool for studying cell surface protein core-fucosylation.

ORGANISM(S): 智人（人类）

SUBMITTER: Xin Guo

PROVIDER: PXD064328 | JPOST Repository | Tue Dec 09 00:00:00 GMT 2025

REPOSITORIES: jPOST

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	GX_QE_easy_20230228_IgG_IGP.raw	Raw
	Hela_O_E_MaxQuant_Result.zip	Other
	Hela_O_E_Raw%20File.zip	Other
	IgG_Experimental%20exploration_MaxQuant_Result.zip	Other
	IgG_Experimental%20exploration_Raw%20File.zip	Other

Items per page:

1 - 5 of 10

Similar Datasets

Project description:Core fucosylation on N-glycosylation plays pivotal roles in regulating ligand binding and cell adhesion, and it is critical to investigate the level of core fucosylation changes in biological processes to further understand its functions. Nevertheless, an unbiased method to globally measure the core fucosylation stoichiometry has not been developed. Here, we devised an approach combining selective enrichment, enzymatic reactions, and multiplexed proteomics to unbiasedly quantify the core fucosylation stoichiometry in multiple biological systems. It was found that the core fucosylation stoichiometry is the lowest in the lysosome and the highest in the extracellular matrix. Moreover, different core fucosylation stoichiometry was observed for the glycosites dwell in various protein domains, and the more aromatic and hydrophobic residues neighboring the glycosites is associated with lower core fucosylation stoichiometry. The method was applied to quantify the core fucosylation stoichiometry change in EMT. It was found that although the overall core fucosylation stoichiometry was similar in the epithelial and mesenchymal states, some glycoproteins involved in extracellular matrix organization and ligand recognition displayed dramatic stoichiometry change, which suggests the regulatory role of core fucosylation in EMT. Breast cell lines (MCF10A, MCF7, and MDA-MB-231) were used as models for cancer progression, and we found that a significant increase of core fucosylation stoichiometry in MDA-MB-231 cells (>80%) comparing to the noncancerous MCF10A cells, but the overall stoichiometry was similar for MCF7 and MCF10A. Moreover, the core fucosylation stoichiometry of an embryonic human kidney cell line, HEK293T was compared with a kidney cancer cell line A498. It was found that the average stoichiometry in HEK293T was the highest among all cell lines quantified in this work (~84%) and was much higher than that of A498 cells, indicating core fucosylation may be an influential regulator in embryonic development. Without any sample restriction, this method can serve as a valuable tool for investigating core fucosylation change in various biological processes.

Dataset Information

Modulation of Core-Fucosylation Oxidation for Selective Enrichment and Characterization of Cell Surface Glycoprotein

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets