title: Structural analyses of O-glycan sugar chains on IgA1 hinge region using SELDI-TOFMS with various lectins.
abstract: The aim of the study was to develop a simple and precise method for identifying glycosylation of the IgA hinge region using surface-enhanced laser desorption/ionization (SELDI)-TOFMS with a lectin-coupled ProteinChip array. Serum IgA was isolated using an anti-IgA antibody column. Following reduction, alkylation, and trypsin digestion, the IgA fragments were applied on the ProteinChip coupled with jacalin, peanut agglutinin (PNA), or Vilsa villosa lectin (VVL). The SELDI-TOFMS peaks corresponding to the fragments containing IgA1 hinge glycopeptides trapped by each lectin were compared. The jacalin-, PNA-, and VVL-immobilized ProteinChips detected 13, 4, and 2 peaks, respectively. One major peak was confirmed as a glycopeptide by MS/MS analysis. These results suggest that a lectin-immobilized ProteinChip assay can be used to simplify the procedures for the analyses of the O-glycans in IgA1 hinge. This method potentially makes it possible to identify a disease-specific glycoform by selecting the appropriate ligand-coupled ProteinChip array.
authors.affiliation: Department of Anatomy, Fujita Health University, School of Medicine, 1-98 Denngakugakubo, Kutsukake-cho, Toyoake, Aichi 470-1192, Japan. kazuo-t@umin.ac.jp
authors.forename: Kazuo
authors.initials: K
authors.lastname: Takahashi
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Hiki
authors.forename: Hiroko
authors.initials: H
authors.lastname: Odani
authors.forename: Sachiko
authors.initials: S
authors.lastname: Shimozato
authors.forename: Hitoo
authors.initials: H
authors.lastname: Iwase
authors.forename: Satoshi
authors.initials: S
authors.lastname: Sugiyama
authors.forename: Nobuteru
authors.initials: N
authors.lastname: Usuda
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002241:Carbohydrates / Q000737:chemistry; D006801:Humans; D007070:Immunoglobulin A / Q000737:chemistry* / Q000276:immunology; D037102:Lectins / Q000737:chemistry* / Q000276:immunology; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry*; D040081:Protein Array Analysis; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D002241:Carbohydrates; D007070:Immunoglobulin A; D037102:Lectins; D011134:Polysaccharides