Abstract
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.
MeSH terms
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Animals
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Cells, Cultured
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Galactosamine / analogs & derivatives
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Galactosamine / chemical synthesis
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Galactosamine / chemistry*
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Humans
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Mice
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Molecular Probes / chemical synthesis
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Molecular Probes / chemistry*
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Molecular Structure
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N-Acetylglucosaminyltransferases / analysis*
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N-Acetylglucosaminyltransferases / metabolism
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beta-N-Acetylhexosaminidases / analysis*
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beta-N-Acetylhexosaminidases / metabolism
Substances
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Molecular Probes
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Galactosamine
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N-Acetylglucosaminyltransferases
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O-GlcNAc transferase
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hexosaminidase C
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beta-N-Acetylhexosaminidases