title: The Small Molecule 2-Azido-2-deoxy-glucose Is a Metabolic Chemical Reporter of O-GlcNAc Modifications in Mammalian Cells, Revealing an Unexpected Promiscuity of O-GlcNAc Transferase.
abstract: Glycans can be directly labeled using unnatural monosaccharide analogs, termed metabolic chemical reporters (MCRs). These compounds enable the secondary visualization and identification of glycoproteins by taking advantage of bioorthogonal reactions. Most widely used MCRs have azides or alkynes at the 2-N-acetyl position but are not selective for one class of glycoprotein over others. To address this limitation, we are exploring additional MCRs that have bioorthogonal functionality at other positions. Here, we report the characterization of 2-azido-2-deoxy-glucose (2AzGlc). We find that 2AzGlc selectively labels intracellular O-GlcNAc modifications, which further supports a somewhat unexpected, structural flexibility in this pathway. In contrast to the endogenous modification N-acetyl-glucosamine (GlcNAc), we find that 2AzGlc is not dynamically removed from protein substrates and that treatment with higher concentrations of per-acetylated 2AzGlc is toxic to cells. Finally, we demonstrate that this toxicity is an inherent property of the small-molecule, as removal of the 6-acetyl-group renders the corresponding reporter nontoxic but still results in protein labeling.
authors.affiliation: Department of Chemistry and ‡Department of Molecular and Computational Biology, University of Southern California , Los Angeles, California 90089-0744, United States.
authors.forename: Balyn W
authors.initials: BW
authors.lastname: Zaro
authors.affiliation: Department of Chemistry and ‡Department of Molecular and Computational Biology, University of Southern California , Los Angeles, California 90089-0744, United States.
authors.forename: Anna R
authors.initials: AR
authors.lastname: Batt
authors.affiliation: Department of Chemistry and ‡Department of Molecular and Computational Biology, University of Southern California , Los Angeles, California 90089-0744, United States.
authors.forename: Kelly N
authors.initials: KN
authors.lastname: Chuh
authors.affiliation: Department of Chemistry and ‡Department of Molecular and Computational Biology, University of Southern California , Los Angeles, California 90089-0744, United States.
authors.forename: Marisol X
authors.initials: MX
authors.lastname: Navarro
authors.affiliation: Department of Chemistry and ‡Department of Molecular and Computational Biology, University of Southern California , Los Angeles, California 90089-0744, United States.
authors.forename: Matthew R
authors.identifier: 0000-0003-3205-5615
authors.initials: MR
authors.lastname: Pratt
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism*; D000818:Animals; D002853:Chromatography, Liquid; D003847:Deoxyglucose / Q000031:analogs & derivatives* / Q000378:metabolism; D051379:Mice; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism*; D041681:NIH 3T3 Cells; D053719:Tandem Mass Spectrometry
chemical_list: D003847:Deoxyglucose; D017351:N-Acetylglucosaminyltransferases; C449880:O-GlcNAc transferase; D000117:Acetylglucosamine