Detection and identification of -GlcNAc-modified proteins using 6-azido-6-deoxy--acetyl-galactosamine

An unnatural monosaccharide with a C6-azide, Ac 3 6AzGalNAc, has been developed as a potent and selective probe for O -GlcNAc-modified proteins. Combined with click chemistry, we demonstrate that Ac 3 6AzGalNAc can robustly label O -GlcNAc glycosylation in a wide range of cell lines. Meanwhile, cell...

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Published in:Organic & biomolecular chemistry 2019-04, Vol.17 (17), p.4326-4334
Main Authors: Guo, Jianshuang, Zhang, Guoqiang, Ma, Jing, Zhao, Caili, Xue, Qingqing, Wang, Jiyan, Liu, Wenjie, Liu, Kaihui, Wang, Haifeng, Liu, Ning, Song, Qitao, Li, Jing
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Summary:An unnatural monosaccharide with a C6-azide, Ac 3 6AzGalNAc, has been developed as a potent and selective probe for O -GlcNAc-modified proteins. Combined with click chemistry, we demonstrate that Ac 3 6AzGalNAc can robustly label O -GlcNAc glycosylation in a wide range of cell lines. Meanwhile, cell imaging and LC-MS/MS proteomics verify its selective activity on O -GlcNAc. More importantly, the protocol presented here provides a general methodology for tracking, capturing and identifying unnatural monosaccharide modified proteins in cells or cell lysates. Ac 3 6AzGalNAc, as a metabolic probe, can robustly label O -GlcNAc-modified proteins in cells with high selectivity.
ISSN:1477-0520
1477-0539
DOI:10.1039/c9ob00516a