Identification of N-glycolylneuraminic acid-containing gangliosides of cat and sheep erythrocytes. 252Cf fission fragment ionization mass spectrometry in the analysis of glycosphingolipids.

K Furukawa,B T Chait,K O Lloyd

doi:10.1016/s0021-9258(18)68129-3

Abstract

A number of gangliosides were isolated from cat and sheep erythrocytes for use in analyzing the specificity of a panel of human anti-heterophile monoclonal antibodies. The structures of these compounds were determined by a combination of different procedures, including sugar analysis, glycosidase treatment, periodate oxidation, TLC immunostaining, methylation analysis, and mass spectrometry. These methods identified the cat erythrocytes gangliosides (C1 and C2) as N-glycolylneuraminic acid (NeuGc)-containing hematosides; C1 was shown to be NeuGc alpha 2----8NeuGc alpha 2----3Gal beta I----4Glc-Cer [NeuGc)2GD3) and C2 to be NeuAc alpha 2----8NeuGc alpha 2----3Gal beta 1----4Glc-Cer [NeuAc-NeuGc-)GD3). The two sheep gangliosides (S1 and S2) were found to be novel glycolipids based on the paragloboside sequence; S1 was identified as NeuGc alpha 2----8NeuGc alpha 2----3Gal beta 1----4GlcNAc beta 1----3Gal beta 1----4Glc-Cer [NeuGc)2-disialylparagloboside) and S2 as NeuAc alpha 2----8NeuGc alpha 2----3Gal beta 1----4GlcNAc beta 1----3Gal beta 1----4Glc-Cer [NeuAc-NeuGc-)-disialylparagloboside). Structural analysis of these compounds was aided by the use of 252Cf fission fragment ionization time-of-flight mass spectrometry. This method provided easily interpretable spectra on methylated derivatives which were particularly useful in determining the sialic acid composition of the gangliosides and the sequence of their disialosyl side chains.

Highlights

A number ofgangliosideswere isolated from cat and side (Z), in bovine erythrocytes N-glycolylneuraminic acidsheep erythrocytesfor usein analyzing the specificity type GM3and NeuGc-sialylparaglobosideare themajor forms of a panel of human anti-heterophile monoclonalnti- [3,4], and erythrocytes of European dogs contain exclusively bodies
The structures tohfese compoundswere deter- (NeuAc)GM8(5).In some animal speciesthe gangliosideshave mined by a combination of different procedures, in- entirely N-acetylneuraminic acid-type (NeuAc)-containing gangliosides (e.g.man) while other tNtcaaoiltnfu-seigaieddlldiyyeoncsstxg;ihoisClde,syaau1lcnntgaideawotueranarmsr,ynaatmaTshshlLsyironCoswsiicpcsny,aeitgccmteltiosydrmgcobao(umeNnsniegeNodtulsraeitGoysuaes.icGin)dTt-icrenchaesogmea2(n,sCt4eetmtaN1himeenynauilentantG,dthgiCpcoohea2ndre)2sim+aoisdda--enssh(-2ipud-eIme3ncs9ia.teMnhsemhaaonpvnrdeoocc3eel2nos-t2sni7rao1elf1lay1na)otnirrabeloaymdcztiioeensdsgtl(ywt6h)iN,ethweseupgGneaoccnit-gfecildocionittsthyiadaionetfsitnwpagorepgosaafennntehgtleliimoonf3GalB1+4Glc-Cer ( ( N ~ u G c ) & ~a~nd) C2 to be non-humananimal erythrocytes, including catand sheep NeuAca2+8NeuGca2+3GalBl+4Glc-Cer
Structural determined the structure of the two cat gangliosides in analysis of these compounds was aided by the use of 2s2Cffission fragment ionization time-of-flight mass spectrometry

Summary

Introduction

A number ofgangliosideswere isolated from cat and side (Z), in bovine erythrocytes N-glycolylneuraminic acidsheep erythrocytesfor usein analyzing the specificity type GM3and NeuGc-sialylparaglobosideare themajor forms of a panel of human anti-heterophile monoclonalnti- [3,4], and erythrocytes of European dogs contain exclusively bodies. Structural determined the structure of the two cat gangliosides in analysis of these compounds was aided by the use of 2s2Cffission fragment ionization time-of-flight mass spectrometry. In the course of studying these structures we utilized 252Cffission position of the gangliosides and the sequence of their fragment ionization mass spectrometry to analyze methylated disialosyl side chains.

Results

Conclusion