title: Glyco-seek: Ultrasensitive Detection of Protein-Specific Glycosylation by Proximity Ligation Polymerase Chain Reaction.
abstract: We report a non-destructive biochemical technique, termed "Glyco-seek", for analysis of O-GlcNAcylated proteins. Glyco-seek combines chemoenzymatic labeling, proximity ligation, and quantitative polymerase chain reaction to detect O-GlcNAcylated proteins with ultrahigh sensitivity. Our glycan-specific assay can be paired with traditional proximity ligation assays to simultaneously determine the change in total protein levels. We show that Glyco-seek detects attomoles of glycoproteins of interest from cell lysates, with sensitivity several orders of magnitude higher than that of current techniques. We used the method to directly assay the O-GlcNAcylation status of a low-abundance transcription factor from cell lysates without need for isolation or enrichment.
authors.affiliation: Departments of †Chemistry, ‡Molecular and Cell Biology, and §Molecular Environmental Biology, University of California , Berkeley, California 94720, United States.|Howard Hughes Medical Institute and ⊥Department of Chemistry, Stanford University , Stanford, California 94305, United States.
authors.forename: Peter V
authors.initials: PV
authors.lastname: Robinson
authors.affiliation: Departments of †Chemistry, ‡Molecular and Cell Biology, and §Molecular Environmental Biology, University of California , Berkeley, California 94720, United States.|Howard Hughes Medical Institute and ⊥Department of Chemistry, Stanford University , Stanford, California 94305, United States.
authors.forename: Cheng-Ting
authors.initials: CT
authors.lastname: Tsai
authors.affiliation: Departments of †Chemistry, ‡Molecular and Cell Biology, and §Molecular Environmental Biology, University of California , Berkeley, California 94720, United States.|Howard Hughes Medical Institute and ⊥Department of Chemistry, Stanford University , Stanford, California 94305, United States.
authors.forename: Amber E
authors.initials: AE
authors.lastname: de Groot
authors.affiliation: Departments of †Chemistry, ‡Molecular and Cell Biology, and §Molecular Environmental Biology, University of California , Berkeley, California 94720, United States.|Howard Hughes Medical Institute and ⊥Department of Chemistry, Stanford University , Stanford, California 94305, United States.
authors.forename: Julia L
authors.initials: JL
authors.lastname: McKechnie
authors.affiliation: Departments of †Chemistry, ‡Molecular and Cell Biology, and §Molecular Environmental Biology, University of California , Berkeley, California 94720, United States.|Howard Hughes Medical Institute and ⊥Department of Chemistry, Stanford University , Stanford, California 94305, United States.
authors.forename: Carolyn R
authors.initials: CR
authors.lastname: Bertozzi
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D001386:Azides / Q000737:chemistry; D057930:Click Chemistry; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D057230:Limit of Detection; D016133:Polymerase Chain Reaction
chemical_list: D001386:Azides; D006023:Glycoproteins; D000117:Acetylglucosamine