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

作者：Xin Guo, Xiaoyan Liu, Zheng Fang, Yan Wang, Lei Liu, Zhongyu Wang, Feng Tang, Wei Huang, Zhenpeng Deng, Qi Wang, Mingliang Ye, Lingyun Jia, Mingming Dong · 发表于：ACS Chemical Biology · 年份：2025 · DOI：10.1021/acschembio.5c00762 · 被引用次数：1 · 研究领域：Glycosylation and Glycoproteins Research、Carbohydrate Chemistry and Synthesis、Infant Nutrition and Health

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, 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-fucosylated 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 investigated the mechanism of core-fucosylation oxidation and developed a new technical tool for studying cell surface protein core-fucos...