Structural analysis of glycosphingolipid analogues obtained by the saccharide primer method using CE-ESI-MS.
A glycosphingolipid analogue (12-azidododecyl beta-lactoside) as a saccharide primer has been shown to be useful for the synthesis of oligosaccharide libraries by mammalian cells. In the present study, CE-ESI-MS was employed to elucidate the structure of glycosphingolipid analogues derived from 12-azidododecyl beta-lactoside (Lac-C12N3) by mammalian cells. MDCK cells and COLO201 cells were cultured with Lac-C12N3, and the glycosylated products secreted into the medium were collected and separated into acidic and neutral products by column chromatography. The acidic products could be directly analyzed by CE-ESI-MS, while the neutral products were converted to anionic derivatives via a reaction with propiolic acid. With this method, it was possible to analyze both acidic and neutral products glycosylated by MDCK cells and COLO201 cells at high sensitivity.
Zhu
Xingyu
X
Department of Biosciences and Informatics, Keio University, Yokohama, Japan.
Hatanaka
Kenichi
K
Yamagata
Tatsuya
T
Sato
Toshinori
T
D000818:Animals; D001386:Azides / Q000737:chemistry*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D019075:Electrophoresis, Capillary / Q000379:methods*; D006028:Glycosphingolipids / Q000096:biosynthesis / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D007785:Lactose / Q000031:analogs & derivatives* / Q000737:chemistry; D009844:Oligosaccharides / Q000138:chemical synthesis*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
C421632:12-azidododecyl lactoside; D001386:Azides; D006028:Glycosphingolipids; D009844:Oligosaccharides; D007785:Lactose