Biochemical studies on sphingolipids of Artemia franciscana: novel neutral glycosphingolipids

Hisao Kojima,Takemasa Shimizu,Mutsumi Sugita,Saki Itonori,Norihisa Fujita,Masahiro Ito

doi:10.1194/jlr.m010173

Abstract

Neutral glycosphingolipids containing one to six sugars in their oligosaccharide chains have been isolated from cysts of the brine shrimp Artemia franciscana. The structures of these glycolipids were identified by methylation analysis, partial acid hydrolysis, gas-liquid chromatography, combined gas-liquid chromatography-mass spectrometry, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and proton nuclear magnetic resonance spectroscopy to be Glcβ1-Cer, Manβ1-4Glcβ1-Cer, Fucα1-3Manβ1-4Glcβ1-Cer, GlcNAcβ1-3Manβ1-4Glcβ1-Cer, GlcNAcα1-2Fucα1-3Manβ1-4Glcβ1-Cer, GalNAcβ1-4GlcNAcβ1-3Manβ1-4Glcβ1-Cer, GalNAcβ1-4(Fucα1-3)GlcNAcβ1-3Manβ1-4Glcβ1-Cer (CPS), and GalNAcβ1-4(GlcNAcα1-2Fucα1-3)GlcNAcβ1-3Manβ1-4Glcβ1-Cer (CHS). Two glycosphingolipids, CPS and CHS, were characterized as novel structures. Because Artemia contains a certain series of glycosphingolipids (-Fucα3Manβ4GlcβCer), which differ from the core sugar sequences reported thus far, we tentatively designated the glycosphingolipids characterized as nonarthro-series ones. Furthermore, CHS exhibited a hybrid structure of arthro-series and nonarthro-series sugar chain. Two novel glycosphingolipids were characterized from the brine shrimp Artemia franciscana; one was composed of arthrotetraose and a branching fucose attached to N-acetylglucosamine residue, and the other was composed of CPS with an additional N-acetylglucosamine residue attached to the branching fucose.

Highlights

Neutral glycosphingolipids containing one to six sugars in their oligosaccharide chains have been isolated from cysts of the brine shrimp Artemia franciscana
Crude sphingolipids (8.6 g) were obtained from chloroform-methanol extracts (285 g) of brine shrimp (Artemia franciscana) cysts (1.8 kg) by alkaline- and acid-treatment and chromatographed on a quaternary ammonium ethyl (QAE)-Sephadex column using a solvent mixture of chloroform-methanol-water (30:60:8, v/v/v)
This study shows the structures of cerebroside, mannolipid, and fucomannolipids as neutral GSLs in the cysts of the brine shrimp Artemia franciscana

Summary

Introduction

Neutral glycosphingolipids containing one to six sugars in their oligosaccharide chains have been isolated from cysts of the brine shrimp Artemia franciscana. The structures of these glycolipids were identified by methylation analysis, partial acid hydrolysis, gas-liquid chromatography, combined gas-liquid chromatography-mass spectrometry, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and proton nuclear magnetic resonance spectroscopy to be Glc␤1-Cer, Man␤1-4Glc␤1-Cer, Fuc␣1-3Man␤1-4Glc␤1-Cer, GlcNAc␤1-3Man␤1-4Glc␤1-Cer, GlcNAc␣1-2Fuc␣1-3Man␤14Glc␤1-Cer, GalNAc␤1-4GlcNAc␤1-3Man␤1-4Glc␤1-Cer, GalNAc␤1-4(Fuc␣1-3)GlcNAc␤1-3Man␤1-4Glc␤1-Cer (CPS), and GalNAc␤1-4(GlcNAc␣1-2Fuc␣1-3)GlcNAc␤1-3Man␤14Glc␤1-Cer (CHS). GSLs, which are expressed on the outer side of the lipid bilayer in animal cells, form patches with sphingomyelin (microdomains) and play a foundational role in intercellular adhesion, cellular recognition, differentiation/growth, and immune response [1,2,3].

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