TY - JOUR
T1 - Liberation of oligosaccharides from glycosphingolipids on PC12 cell surface with endoglycoceramidase
AU - Rasilo, Maija Liisa
AU - Ito, Makoto
AU - Yamagata, Tatsuya
N1 - Funding Information:
ACKNOWLEDGMENWTSe: are indebted to Dr. T. Amano for the supply of PC12 cells, to Drs. T. Ikekita and C. C. Sweeley for their help in the derivatization of oligosaccharides with ABEE, and to Drs. Higashi and l4. Shimamura for their helpful comments. We are grateful to I. Hosokane and Y. Ikegami for their technical assistance. This work was supported in part by the Cultural Foundation of Finland and the Emil Aaltonen Foundation.
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1989/8/15
Y1 - 1989/8/15
N2 - Endoglycoceramidase (EGCase) cleaves the linkage between the oligosaccharide and ceramide of glycosphingolipids (Ito, M., and Yamagata, T., J. Biol. Chem. 261, 14278-14282, ibid, 264, in press). Intact cells of rat pheochromocytoma line PC12 were treated with the highly purified EGCase I and the oligosaccharides released were analyzed by HPLC. Cleavage of the oligosaccharides with the enzyme reached a plateau as the amount of the enzyme was increased. At maximum, 42% of the oligosaccharides from globoside, 40% from GalGb3Cer, and 60% from Gb3Cer were liberated during 1h of incubation without impairing the viability of cells. The only partial liberation indicates that not all oligosaccharides of cell surface glycosphingolipids are accessible to the enzyme. The use of EGCase offers an important new method to access the functions of glycosphingolipids on cell surface in situ.
AB - Endoglycoceramidase (EGCase) cleaves the linkage between the oligosaccharide and ceramide of glycosphingolipids (Ito, M., and Yamagata, T., J. Biol. Chem. 261, 14278-14282, ibid, 264, in press). Intact cells of rat pheochromocytoma line PC12 were treated with the highly purified EGCase I and the oligosaccharides released were analyzed by HPLC. Cleavage of the oligosaccharides with the enzyme reached a plateau as the amount of the enzyme was increased. At maximum, 42% of the oligosaccharides from globoside, 40% from GalGb3Cer, and 60% from Gb3Cer were liberated during 1h of incubation without impairing the viability of cells. The only partial liberation indicates that not all oligosaccharides of cell surface glycosphingolipids are accessible to the enzyme. The use of EGCase offers an important new method to access the functions of glycosphingolipids on cell surface in situ.
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U2 - 10.1016/0006-291X(89)90785-7
DO - 10.1016/0006-291X(89)90785-7
M3 - Article
C2 - 2764919
AN - SCOPUS:0024365996
SN - 0006-291X
VL - 162
SP - 1093
EP - 1099
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -