Metabolic cross-talk allows labeling of O-linked beta-N-acetylglucosamine-modified proteins via the N-acetylgalactosamine salvage pathway.
| dc.contributor.author | Boyce, Michael | |
| dc.contributor.author | Carrico, Isaac S | |
| dc.contributor.author | Ganguli, Anjali S | |
| dc.contributor.author | Yu, Seok-Ho | |
| dc.contributor.author | Hangauer, Matthew J | |
| dc.contributor.author | Hubbard, Sarah C | |
| dc.contributor.author | Kohler, Jennifer J | |
| dc.contributor.author | Bertozzi, Carolyn R | |
| dc.date.accessioned | 2020-01-01T17:04:10Z | |
| dc.date.available | 2020-01-01T17:04:10Z | |
| dc.date.issued | 2011-02-07 | |
| dc.date.updated | 2020-01-01T17:04:09Z | |
| dc.description.abstract | Hundreds of mammalian nuclear and cytoplasmic proteins are reversibly glycosylated by O-linked β-N-acetylglucosamine (O-GlcNAc) to regulate their function, localization, and stability. Despite its broad functional significance, the dynamic and posttranslational nature of O-GlcNAc signaling makes it challenging to study using traditional molecular and cell biological techniques alone. Here, we report that metabolic cross-talk between the N-acetylgalactosamine salvage and O-GlcNAcylation pathways can be exploited for the tagging and identification of O-GlcNAcylated proteins. We found that N-azidoacetylgalactosamine (GalNAz) is converted by endogenous mammalian biosynthetic enzymes to UDP-GalNAz and then epimerized to UDP-N-azidoacetylglucosamine (GlcNAz). O-GlcNAc transferase accepts UDP-GlcNAz as a nucleotide-sugar donor, appending an azidosugar onto its native substrates, which can then be detected by covalent labeling using azide-reactive chemical probes. In a proof-of-principle proteomics experiment, we used metabolic GalNAz labeling of human cells and a bioorthogonal chemical probe to affinity-purify and identify numerous O-GlcNAcylated proteins. Our work provides a blueprint for a wide variety of future chemical approaches to identify, visualize, and characterize dynamic O-GlcNAc signaling. | |
| dc.identifier | 1010045108 | |
| dc.identifier.issn | 0027-8424 | |
| dc.identifier.issn | 1091-6490 | |
| dc.identifier.uri | ||
| dc.language | eng | |
| dc.publisher | Proceedings of the National Academy of Sciences | |
| dc.relation.ispartof | Proceedings of the National Academy of Sciences of the United States of America | |
| dc.relation.isversionof | 10.1073/pnas.1010045108 | |
| dc.subject | Cell Line | |
| dc.subject | Humans | |
| dc.subject | Acetylgalactosamine | |
| dc.subject | Acetylglucosamine | |
| dc.subject | Affinity Labels | |
| dc.subject | Chromatography, Affinity | |
| dc.subject | Methods | |
| dc.subject | Receptor Cross-Talk | |
| dc.subject | Protein Processing, Post-Translational | |
| dc.subject | Glycosylation | |
| dc.subject | Metabolic Networks and Pathways | |
| dc.title | Metabolic cross-talk allows labeling of O-linked beta-N-acetylglucosamine-modified proteins via the N-acetylgalactosamine salvage pathway. | |
| dc.type | Journal article | |
| duke.contributor.orcid | Boyce, Michael|0000-0002-2729-4876 | |
| pubs.begin-page | 3141 | |
| pubs.end-page | 3146 | |
| pubs.issue | 8 | |
| pubs.organisational-group | School of Medicine | |
| pubs.organisational-group | Duke | |
| pubs.organisational-group | Duke Cancer Institute | |
| pubs.organisational-group | Institutes and Centers | |
| pubs.organisational-group | Biochemistry | |
| pubs.organisational-group | Basic Science Departments | |
| pubs.publication-status | Published | |
| pubs.volume | 108 |
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