Unbiased quantification of core-fucosylation stoichiometry reveals its role in cancer progression and embryonic development

ABSTRACT: 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.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Ronghu Wu  

PROVIDER: MSV000093936 | MassIVE | Fri Jan 26 13:17:00 GMT 2024

REPOSITORIES: MassIVE