title: Detection and analysis of proteins modified by O-linked N-acetylglucosamine.
abstract: First, a protocol for increasing the stoichiometry of O-GlcNAc on proteins is given. This is followed by simple techniques for the detection/screening of O-GlcNAc-modified proteins either by immunoblotting or lectin affinity chromatography. Separate protocols verify that the glycan is O-linked GlcNAc. These methods are followed by protocols for more comprehensive analysis of O-GlcNAc modified proteins, including labeling of O-GlcNAc residues with [3H]Gal, and subsequent product analysis. The final two protocols assay for O-GlcNAc transferase and O-GlcNAcase activity, respectively.
authors.affiliation: The Johns Hopkins University, Medical School, Baltimore, Maryland, USA.
authors.forename: N E
authors.initials: NE
authors.lastname: Zachara
authors.forename: R N
authors.initials: RN
authors.lastname: Cole
authors.forename: G W
authors.initials: GW
authors.lastname: Hart
authors.forename: Y
authors.initials: Y
authors.lastname: Gao
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry*; D002846:Chromatography, Affinity; D015151:Immunoblotting; D037102:Lectins / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D037102:Lectins; D011506:Proteins; D000117:Acetylglucosamine