Characterization of a major neutral glycolipid in PC12 cells as III3Galα-globotriaosylceramide by the method for determining glycosphingolipid saccharide sequence with endoglycoceramidase

M. Shimamura, T. Hayase, M. Ito, M. L. Rasilo, T. Yamagata

研究成果: Contribution to journalArticle査読

22 被引用数 (Scopus)

抄録

Neutral glycolipids in PC12 cells were examined. A major neutral glycosphingolipid, isolated from a chloroform/methanol extract of the cells, was found to contain only galactose and glucose at a ratio of 3:1 and identified as ceramide tetrahexoside by fast atom bombardment (FAB) mass spectrometry. Its saccharide sequence was determined by a new method developed here using endoglycoceramidase (Ito, M., and Yamagata, T. (1986) J. Biol. Chem. 261, 14278-14282). The glycosphingolipid was digested with endoglycoceramidase to produce oligosaccharide which was subsequently pyridylaminated. The fluorescence-labeled oligosaccharide was digested with a series of specific exoglycosidases and fractionated by high performance liquid chromatography. The 2-aminopyridyl oligosaccharide was hydrolyzed by α-galactosidase to give a 2-aminopyridyl oligosaccharide which was identified as 2-aminopyridyl lactose by high performance liquid chromatography, indicating the glycolipid structure to be GalαGalαGalβGlcCer. Ceramide trihexoside obtained by limited digestion of the intact glycolipid was clearly identical with ceramide trihexoside obtained from human erythrocytes, according to NMR spectroscopy and methylation analysis. From these and other data on the intact glycolipid, obtained by methylation analysis and NMR spectroscopy, its structure was confirmed as Galα1-3Galα1-4Galβ1-4Glcβ1-1Cer, III3-Galα-globotriaosylceramide. This is the first report indicating the presence of this glycosphingolipid in PC12 cells.

本文言語英語
ページ(範囲)12124-12128
ページ数5
ジャーナルJournal of Biological Chemistry
263
24
出版ステータス出版済み - 1988

All Science Journal Classification (ASJC) codes

  • 生化学
  • 分子生物学
  • 細胞生物学

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