title: Global identification of O-GlcNAc-modified proteins.
abstract: The O-linked N-acetylglucosamine (O-GlcNAc) modification of serine/threonine residues is an abundant posttranslational modification present in cytosolic and nuclear proteins. The functions and subproteome of O-GlcNAc modification remain largely undefined. Here we report the application of the tagging-via-substrate (TAS) approach for global identification of O-GlcNAc-modified proteins. The TAS method utilizes an O-GlcNAc azide analogue for metabolic labeling of O-GlcNAc-modified proteins, which can be chemoselectively conjugated for detection and enrichment of the proteins for proteomics studies. Our study led to the identification of 199 putative O-GlcNAc-modified proteins from HeLa cells, among which 23 were confirmed using reciprocal immunoprecipitation. Functional classification shows that proteins with diverse functions are modified by O-GlcNAc, implying that O-GlcNAc might be involved in the regulation of multiple cellular pathways.
authors.affiliation: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9038, USA.
authors.forename: Animesh
authors.initials: A
authors.lastname: Nandi
authors.forename: Robert
authors.initials: R
authors.lastname: Sprung
authors.forename: Deb K
authors.initials: DK
authors.lastname: Barma
authors.forename: Yingxin
authors.initials: Y
authors.lastname: Zhao
authors.forename: Sung Chan
authors.initials: SC
authors.lastname: Kim
authors.forename: John R
authors.initials: JR
authors.lastname: Falck
authors.forename: Yingming
authors.initials: Y
authors.lastname: Zhao
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D006367:HeLa Cells; D006801:Humans; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D011506:Proteins; D017351:N-Acetylglucosaminyltransferases; D000117:Acetylglucosamine