title: Detection and identification of O-GlcNAcylated proteins by proteomic approaches.
abstract: O-GlcNAcylation (O-linked beta-N-acetylglucosaminylation) is a widespread PTM confined within the nuclear, the cytosolic, and the mitochondrial compartments of eukaryotes. Recently, O-GlcNAcylation has been also detected in the close vicinity of plasma membranes particularly in lipid microdomains. The detection of this PTM can be easily done if appropriate controls and precautions are taken using a wide variety of tools including lectins, antibodies, or click-chemistry-based methods. In contrast, the identification of the proteins bearing O-GlcNAc moieties and the localization of the precise sites of O-GlcNAcylation remain challenging. This is due to the lability of the glycosidic bond between hydroxyl group of serine or threonine and N-acetylglucosamine using conventional fragmentation techniques such as CID. To tentatively overcome this technical limitation, electron-capture dissociation, or electron-transfer dissociation MS/MS are now used. Thanks to these breakthroughs, a large number of O-GlcNAc sites have been identified to date but these methodologies remain far from being used in routine.
authors.affiliation: Unit of Structural and Functional Glycobiology, University of Lille 1, Villeneuve d'Ascq, France.
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Vercoutter-Edouart
authors.forename: Ikram
authors.initials: I
authors.lastname: El Yazidi-Belkoura
authors.forename: Céline
authors.initials: C
authors.lastname: Guinez
authors.forename: Steffi
authors.initials: S
authors.lastname: Baldini
authors.forename: Maïté
authors.initials: M
authors.lastname: Leturcq
authors.forename: Marlène
authors.initials: M
authors.lastname: Mortuaire
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Mir
authors.forename: Agata
authors.initials: A
authors.lastname: Steenackers
authors.forename: Vanessa
authors.initials: V
authors.lastname: Dehennaut
authors.forename: Annick
authors.initials: A
authors.lastname: Pierce
authors.forename: Tony
authors.initials: T
authors.lastname: Lefebvre
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D051379:Mice; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D051381:Rats; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D006023:Glycoproteins; D000117:Acetylglucosamine
keywords: Glycoproteomics; Mass Spectrometry; O-GlcNAcome; O-GlcNAcylation; Post-translational modification; Site mapping