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

An unnatural monosaccharide with a C6-azide, Ac36AzGalNAc, has been developed as a potent and selective probe for O-GlcNAc-modified proteins. Combined with click chemistry, we demonstrate that Ac36AzGalNAc can robustly label O-GlcNAc glycosylation in a wide range of cell lines. Meanwhile, cell imagi...

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Bibliographic Details
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
Format: Article
Language:English
Subjects:
Animals
beta-N-Acetylhexosaminidases - analysis
beta-N-Acetylhexosaminidases - metabolism
Cell lines
Cells, Cultured
Chemical synthesis
D-Galactosamine
Galactosamine - analogs & derivatives
Galactosamine - chemical synthesis
Galactosamine - chemistry
Glycosylation
Humans
Lysates
Mice
Molecular Probes - chemical synthesis
Molecular Probes - chemistry
Molecular Structure
Monosaccharides
N-Acetylgalactosamine
N-Acetylglucosaminyltransferases - analysis
N-Acetylglucosaminyltransferases - metabolism
Organic chemistry
Proteins
Proteomics
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Description
Summary:An unnatural monosaccharide with a C6-azide, Ac36AzGalNAc, has been developed as a potent and selective probe for O-GlcNAc-modified proteins. Combined with click chemistry, we demonstrate that Ac36AzGalNAc 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.
ISSN:1477-0520
1477-0539
DOI:10.1039/c9ob00516a