Title:  Chemoenzymatic Labeling, Detection and Profiling of Core Fucosylation in Live Cells

Qiang Zhu, Jean-Luc Chaubard, Didi Geng, Jiechen Shen, Lan Ban, Sheldon T. Cheung, Fangyu Wei, Yating Liu, Haofan Sun, Angie Calderon, Wenbo Dong, Weijie Qin, Tiehai Li, Liuqing Wen, Peng George Wang, Shisheng Sun, Wen Yi, and Linda C. Hsieh-Wilson

Abstract

Core fucosylation, the attachment of an α-1,6-linked-fucose to the N-glycan core pentasaccharide, is an abundant protein modification that plays critical roles in various biological processes such as cell signaling, B cell development, antibody-dependent cellular cytotoxicity, and oncogenesis. However, the tools currently used to detect core fucosylation suffer from poor specificity, exhibiting cross-reactivity against all types of fucosylation. Herein we report the development of a new chemoenzymatic strategy for the rapid and selective detection of core fucosylated glycans. This approach employs a galactosyltransferase enzyme identified from Caenorhabditis elegans that specifically transfers an azido-appended galactose residue onto core fucose via a β-1,4 glycosidic linkage. We demonstrate that the approach exhibits superior specificity toward core fucose on a variety of complex N-glycans. The method enables detection of core fucosylated glycoproteins from complex cell lysates, as well as on live cell surfaces, and it can be integrated into a diagnostic platform to profile protein-specific core fucosylation levels. This chemoenzymatic labeling approach offers a new strategy for the identification of disease biomarkers and will allow researchers to further characterize the fundamental role of this important glycan in normal and disease physiology.

Link: https://pubs.acs.org/doi/10.1021/jacs.4c09303