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

Area of Science:

• Glycobiology
• Proteomics
• Biochemistry

Background:

• Protein core-fucosylation (CF) is vital for cell surface protein function, impacting cell signaling, immune response, and cancer.
• Identifying and characterizing CF is challenging due to low abundance and N-glycosylation heterogeneity.

Purpose of the Study:

• To investigate core-fucose oxidation mechanisms.
• To develop a selective enrichment strategy for cell surface core-fucosylated glycopeptides.
• To establish a new tool for characterizing cell surface protein core-fucosylation.

Main Methods:

• Selective oxidation of cell surface glycans using sodium periodate.
• Enzymatic truncation of N-glycans with Endo M and Endo F3.
• Selective enrichment of core-fucosylated glycopeptides using reversible hydrazide chemistry.

Main Results:

• Developed a strategy combining chemical oxidation and enzymatic truncation for CF characterization.
• Successfully profiled cell surface core-fucosylated proteins in HeLa cells.
• Identified 74 core-fucosylated glycopeptides from 21 cell surface drug targets.

Conclusions:

• The developed method effectively overcomes challenges in identifying and characterizing cell surface core-fucosylation.
• This approach provides a valuable tool for studying the biological roles of core-fucosylated proteins.
• The findings validate the strategy's efficacy in profiling key cell surface drug targets.