Structural analyses of O-glycan sugar chains on IgA1 hinge region using SELDI-TOFMS with various lectins
- 作者：Kazuo Takahashi ; Yoshiyuki Hiki ; Hiroko Odani ; Sachiko Shimozato ; Hitoo Iwase ; Satoshi Sugiyama and Nobuteru Usuda
- 关键词：O-Glycan ; O-Glycosylation ; N-Acetylgalactosamine ; Galactose ; Sialic acid ; IgA1 hinge peptide ; Lectin assay ; SELDI-TOFMS ; ESI/ITMS
- 刊名：Biochemical and Biophysical Research Communications
- 出版年：2006
- 期刊代码：159_0006291x
- 类别：bio
- 出版时间：24 November 2006
- 卷：350
- 期：3
- 页码：580-587
- 文件大小：593 K
摘要
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.