Isolation and characterization of gangliosides from Theileria sergenti

Abstract

The gangliosides of Theileria sergenti piroplasms were isolated and analyzed by thin-layer chromatography (TLC) and TLC immunostaining test. Four species of gangliosides, designated as G-1, G-2, G-3, and G-4, were separated on TLC. G-1, G-2, G-3, and G-4 ganglioside showed the same mobility as GM3, sialosylparagloboside (SPG), i-active ganglioside, and I-active ganglioside on the TLC plate, respectively. In order to characterize the molecular species of gangliosides from T. sergenti, G-1, G-2, G-3, and G-4 gangliosides were purified and tested by TLC immunostaining test with monoclonal antibodies against gangliosides. G-1 ganglioside had reactivity to anti-GM3 monoclonal antibody. G-2 gave reaction with monoclonal antibody to SPG containing N-glycolylneuraminic acid (NeuGc). G-3 showed reactivity to the anti-i-active ganglioside (NeuGc) monoclonal antibody. G-4 was recognized by the monoclonal antibody which reacts with I-active ganglioside (NeuGc). In addition, sialic acid moiety of the gangliosides from T. sergenti piroplasms was also analyzed. N-acetylneuraminic acid-containing gangliosides were hardly detectable in T. sergenti piroplasms. Gangliosides from T. sergenti (G-1, G-2, G-3, and G-4) carried only NeuGc as their sialic acid moiety. These results suggest that G-1, G-2, G-3, and G-4 gangliosides are GM3 (NeuGc) [NeuGcα2-3Galβ1-4Glcβ1-1Cer], SPG (NeuGc) [NeuGcα2-3Gal/β1-4GlcNAcβ1-3Galβ1-4Glcβ1-1Cer], i-active ganglioside (NeuGc) [NeuGcα2-3Galβ1-4GlcNAcβ1-3Gaβ1-4GlcNAcβ1-3Galβ1- 4Glcββ1-1Cer], and I-active ganglioside(NeuGc) [NeuGcα2-3Galβ1-4GlcNAcβ1-3 (Galα1-3Galβ1-4GlcNAcβ1-6) Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1Cer], respectively.