Detection and identification of O -GlcNAc-modified proteins using 6-azido-6-deoxy- N -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 Vol. 17; no. 17; pp. 4326 - 4334 |
|---|---|
| Main Authors | , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
England
Royal Society of Chemistry
24.04.2019
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| Subjects | |
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| Abstract | 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. |
|---|---|
| AbstractList | 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. 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.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. 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. |
| Author | Zhao, Caili Ma, Jing Liu, Ning Song, Qitao Guo, Jianshuang Zhang, Guoqiang Li, Jing Xue, Qingqing Liu, Wenjie Liu, Kaihui Wang, Jiyan Wang, Haifeng |
| Author_xml | – sequence: 1 givenname: Jianshuang surname: Guo fullname: Guo, Jianshuang organization: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China – sequence: 2 givenname: Guoqiang surname: Zhang fullname: Zhang, Guoqiang organization: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China – sequence: 3 givenname: Jing surname: Ma fullname: Ma, Jing organization: Institute of Chemical Biology, College of Pharmacy, Henan University, Kaifeng 475004, China – sequence: 4 givenname: Caili surname: Zhao fullname: Zhao, Caili organization: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China – sequence: 5 givenname: Qingqing surname: Xue fullname: Xue, Qingqing organization: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China – sequence: 6 givenname: Jiyan surname: Wang fullname: Wang, Jiyan organization: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China – sequence: 7 givenname: Wenjie surname: Liu fullname: Liu, Wenjie organization: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China – sequence: 8 givenname: Kaihui surname: Liu fullname: Liu, Kaihui organization: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China – sequence: 9 givenname: Haifeng surname: Wang fullname: Wang, Haifeng organization: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China – sequence: 10 givenname: Ning surname: Liu fullname: Liu, Ning organization: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China – sequence: 11 givenname: Qitao surname: Song fullname: Song, Qitao organization: College of Chemistry and Molecular Engineering, Beijing National Laboratory for Molecular Sciences, Peking University, Beijing 100191, China – sequence: 12 givenname: Jing orcidid: 0000-0002-4289-4829 surname: Li fullname: Li, Jing organization: State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300353, China |
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| Snippet | 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... An unnatural monosaccharide with a C6-azide, Ac36AzGalNAc, has been developed as a potent and selective probe for O-GlcNAc-modified proteins. Combined with... |
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| SubjectTerms | 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 |
| Title | Detection and identification of O -GlcNAc-modified proteins using 6-azido-6-deoxy- N -acetyl-galactosamine |
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