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 inOrganic & biomolecular chemistry Vol. 17; no. 17; pp. 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
Format Journal Article
Published 24.04.2019
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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.
Bibliography:Electronic supplementary information (ESI) available. See DOI
10.1039/c9ob00516a
ISSN:1477-0520
1477-0539
DOI:10.1039/c9ob00516a