title: Cellular O-Glycome Reporter/Amplification to explore O-glycans of living cells.
abstract: Protein O-glycosylation has key roles in many biological processes, but the repertoire of O-glycans synthesized by cells is difficult to determine. Here we describe an approach termed Cellular O-Glycome Reporter/Amplification (CORA), a sensitive method used to amplify and profile mucin-type O-glycans synthesized by living cells. Cells convert added peracetylated benzyl-α-N-acetylgalactosamine to a large variety of modified O-glycan derivatives that are secreted from cells, allowing for easy purification for analysis by HPLC and mass spectrometry (MS). Relative to conventional O-glycan analyses, CORA resulted in an ∼100-1,000-fold increase in sensitivity and identified a more complex repertoire of O-glycans in more than a dozen cell types from Homo sapiens and Mus musculus. Furthermore, when coupled with computational modeling, CORA can be used for predictions about the diversity of the human O-glycome and offers new opportunities to identify novel glycan biomarkers for human diseases.
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA.|Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Kudelka
authors.affiliation: Department of Life Sciences, Imperial College London, London, UK.
authors.forename: Aristotelis
authors.initials: A
authors.lastname: Antonopoulos
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Yingchun
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Duc M
authors.initials: DM
authors.lastname: Duong
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Xuezheng
authors.initials: X
authors.lastname: Song
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Nicholas T
authors.initials: NT
authors.lastname: Seyfried
authors.affiliation: Department of Life Sciences, Imperial College London, London, UK.
authors.forename: Anne
authors.initials: A
authors.lastname: Dell
authors.affiliation: Department of Life Sciences, Imperial College London, London, UK.
authors.forename: Stuart M
authors.initials: SM
authors.lastname: Haslam
authors.affiliation: Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Richard D
authors.initials: RD
authors.lastname: Cummings
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Tongzhong
authors.initials: T
authors.lastname: Ju
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D013058:Mass Spectrometry; D051379:Mice; D011134:Polysaccharides / Q000378:metabolism*
chemical_list: D011134:Polysaccharides