Preparation of homogenous oligosaccharide chains from glycosphingolipids.
After the discovery of glycosphingolipid (GSL) glycan detaching enzymes, Rhodococcal endoglycoceramidase (EGCase) and leech ceramide glycanase (CGase), the method for enzymatically releasing glycans from GSLs has become the method of choice for preparing intact ceramide-free oligosaccharide chains from GSLs. This paper describes (1) the preparation of the intact oligosaccharides from GM1 (II(3)NeuAcGgOse(4)Cer) and GbOse(4)Cer as examples to show the use of CGase to prepare intact glycan chains from GSLs, and (2) the specificity and detergent requirements of Rhodococcal EGCases for the release of glycan chains from different GSLs.
Li
Yu-Teh
YT
Department of Biochemistry, Tulane University Health Sciences Center School of Medicine, New Orleans, LA 70112, USA. yli1@tulane.edu
Chou
Chau-Wen
CW
Li
Su-Chen
SC
Kobayashi
Utaro
U
Ishibashi
Yo-hey
YH
Ito
Makoto
M
D000818:Animals; D002240:Carbohydrate Sequence; D002855:Chromatography, Thin Layer; D003902:Detergents / Q000494:pharmacology; D005677:G(M1) Ganglioside / Q000032:analysis; D005915:Globosides / Q000032:analysis / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000378:metabolism; D006028:Glycosphingolipids / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis / Q000187:drug effects; D007865:Leeches; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry; D012240:Rhodococcus / Q000201:enzymology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity / Q000187:drug effects
D003902:Detergents; D005915:Globosides; D006028:Glycosphingolipids; D009844:Oligosaccharides; C024032:globotetraosylceramide; D005677:G(M1) Ganglioside; D006026:Glycoside Hydrolases; C050427:endoglycoceramidase