A novel fucosyl glycosphingolipid of brine shrimp that is highly sensitive to endoglycoceramidase.
Endoglycoceramidase (EGCase; EC 3.2.1.123) is a glycohydrolase that hydrolyzes the glycosidic linkage between the oligosaccharide and ceramide of various glycosphingolipids. We previously reported that hydra produced EGCase to digest glycosphingolipids of brine shrimp (Artemia salina), a type of aquatic crustacean used as a diet for the culture of hydra (Horibata Y, Sakaguchi K, Okino N, Iida H, Inagaki M, Fujisawa T, Hama Y, Ito M. 2004. J Biol Chem. 279:33379-33389). We report here that a major glycosphingolipid of brine shrimp is unique in structure and highly sensitive to EGCase. The glycosphingolipid was extracted from freshly hatched brine shrimp by Folch's partition, followed by mild alkaline hydrolysis and purification with a Sep-Pak plus silica cartridge. The structure of brine shrimp glycosphingolipid was determined by gas chromatography, gas chromatography-mass spectrometry, fast-atom bombardment mass spectrometry, and (1)H-NMR spectrometry to be GlcNAcalpha1-2Fucalpha1-3Manbeta1-4Glcbeta1-1'Cer. Two major molecular species of the glycosphingolipid were identified; the sugar and sphingoid base of each were the same but the major fatty acid was C22:0 and 2-hydroxy C22:0, respectively. This is the first report describing the glycosphingolipid that has an internal fucosyl residue substituted with alpha1-2 N-acetylglucosaminyl residue. This study also suggests the biological relevance of the glycosphingolipid as a dietary source of hydra which possesses EGCase as a digestion enzyme.
Xu
Xu
X
Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan.
Horibata
Yasuhiro
Y
Inagaki
Masanori
M
Hama
Yoichiro
Y
Sakaguchi
Keishi
K
Goda
Hatsumi M
HM
Okino
Nozomu
N
Ito
Makoto
M
D000818:Animals; D001156:Artemia / Q000737:chemistry / Q000378:metabolism*; D002240:Carbohydrate Sequence; D006017:Glycolipids / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006028:Glycosphingolipids / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D006868:Hydrolysis; D008954:Models, Biological; D008968:Molecular Conformation; D013379:Substrate Specificity
D006017:Glycolipids; D006028:Glycosphingolipids; C007790:fucosylglycolipids; D006026:Glycoside Hydrolases; C050427:endoglycoceramidase