title: Glycosylation sites identified by solid-phase Edman degradation: O-linked glycosylation motifs on human glycophorin A.
abstract: The human red blood cell sialoglycoprotein, glycophorin A (GpA), contains a 'mucin-like' extensively O-glycosylated extracellular domain which carries the MN blood group antigens. We have revised the sites of O-glycosylation in the extracellular domain of GpA by automated solid-phase Edman degradation, which allowed positive identification and quantitation of O-glycosylated Ser and Thr residues, as well as the single N-glycosylation site. One N-linked and 16 O-linked sites were identified. Carbohydrate was absent on Ser1, Ser14, Ser15, Ser23, Thr28 and Thr58 in GpA. We propose that the glycosyltransferases present in erythrocytes recognize specific flanking sequences around potential O-glycosylation sites. All 16 O-glycosylation sites are explained on the basis of four motifs. Three motifs are associated with Thr-glycosylation: Xaa-Pro-Xaa-Xaa where at least one Xaa = Thr; Thr-Xaa-Xaa-Xaa where at least one Xaa = Thr; Xaa-Xaa-Thr-Xaa where at least one X = Arg or Lys. The fourth motif is associated with Ser-glycosylation: Ser-Xaa-Xaa-Xaa where at least one Xaa = Ser. These simple rules explain the glycosylation (or lack of it) on 21 of 22 Ser/Thr in the extracellular domain of GpA.
authors.affiliation: Macquarie University Centre for Analytical Biotechnology, Macquarie, NSW, Australia.
authors.forename: A
authors.initials: A
authors.lastname: Pisano
authors.forename: J W
authors.initials: JW
authors.lastname: Redmond
authors.forename: K L
authors.initials: KL
authors.lastname: Williams
authors.forename: A A
authors.initials: AA
authors.lastname: Gooley
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D006021:Glycophorins / Q000737:chemistry* / Q000235:genetics; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D014357:Trypsin
chemical_list: D006021:Glycophorins; D010446:Peptide Fragments; D014357:Trypsin