Glycobiology Advance Access originally published online on August 21, 2009
Glycobiology 2009 19(12):1446-1451; doi:10.1093/glycob/cwp118
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A novel fucosyl glycosphingolipid of brine shrimp that is highly sensitive to endoglycoceramidase
2 Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan
3 College of Life Science, Shenzhen University, Nanhai Ave 3688, Shenzhen 518060, China
4 Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582
5 Faculty of Agriculture, Saga University, 1 Honjo, Saga 840-8502
6 Bio-architecture Center, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan
1 To whom correspondence should be addressed: Tel: +81-92-642-2898; Fax: +81-92-642-2907; e-mail: makotoi{at}agr.kyushu-u.ac.jp
Received on June 4, 2009; revised on August 7, 2009; accepted on August 9, 2009
Endoglycoceramidase (EGCase; EC 3.2.1.123
[EC]
) 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 1H-NMR spectrometry to be GlcNAc
1-2Fuc1-3Manβ1-4Glcβ1-1'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 1-2 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.
Key words: brine shrimp / endoglycoceramidase / fucosyl / glycosphingolipid