title: The glycosylation of the complement regulatory protein, human erythrocyte CD59.
abstract: Human erythrocyte CD59 contains N- and O-glycans and a glycosylphosphatidylinositol (GPI) anchor, all of which have been analyzed in this study. The anchor consists principally of the minimum core glycan sequence Manalpha1-2Manalpha1-6Manalpha1-4GlcN-linked to a phosphatidylinositol moiety with the structure sn-1-O-alkyl(C18:0 and C18:1)-2-O-acyl(C20:4)glycerol-3-phospho-1-(2-O-palmitoyl(C16:0))myo- inositol. This structure is essentially identical to that of human erythrocyte cholinesterase (Deeg, M. A., Humphrey, D. R., Yang, S. H. , Ferguson, T. R., Reinhold, V. N., and Rosenberry, T. L. (1992) J. Biol. Chem. 267, 18573-18580). This first comparison of GPI anchors from different proteins expressed in the same tissue suggests that human reticulocytes produce only one type of anchor structure. The N- and O-glycans were sequenced using a novel approach involving digestion of the total glycan pool with multiple enzyme arrays. The N-glycan pool contained families of bi-antennary complex-type structures with and without lactosamine extensions and outer arm fucose residues. The predominant O-glycans were NeuNAcalpha2-3Galbeta1-3GalNAc and Galbeta1-3[NeuNAcalpha2-3]GalNAc. Inspection of a molecular model of CD59, based on the NMR solution structure of the extracellular domain and the structural data from this study, suggested several roles for the glycans, including spacing and orienting CD59 on the cell surface and protecting the molecule from proteases. This work completes the initial structural analysis of CD59, providing the most complete view of any cell surface glycoprotein studied to date.
authors.affiliation: Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, United Kingdom.
authors.forename: P M
authors.initials: PM
authors.lastname: Rudd
authors.forename: B P
authors.initials: BP
authors.lastname: Morgan
authors.forename: M R
authors.initials: MR
authors.lastname: Wormald
authors.forename: D J
authors.initials: DJ
authors.lastname: Harvey
authors.forename: C W
authors.initials: CW
authors.lastname: van den Berg
authors.forename: S J
authors.initials: SJ
authors.lastname: Davis
authors.forename: M A
authors.initials: MA
authors.lastname: Ferguson
authors.forename: R A
authors.initials: RA
authors.lastname: Dwek
mesh_terms: D018957:CD59 Antigens / Q000032:analysis / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003169:Complement Inactivator Proteins / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D008958:Models, Molecular
chemical_list: D018957:CD59 Antigens; D003169:Complement Inactivator Proteins