ISOLATION AND CHARACTERIZATION OF GANGLIOSIDES FROM THEILERIA-SERGENTI

The gangliosides of Theileria sergenti piroplasms were isolated and an alyzed by thin-layer chromatography (TLC) and TLC immunostaining test. Four species of gangliosides, designated as G-1, G-2, G-3, and G-4, w ere separated on TLC. G-1, G-2, G-3, and G-4 ganglioside showed the sa me 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 imm unostaining test with monoclonal antibodies against gangliosides. G-1 ganglioside had reactivity to anti-GM3 monoclonal antibody. G-2 gave r eaction with monoclonal antibody to SPG containing N-glycolylneuramini c acid (NeuGc). G-3 showed reactivity to the anti-i-active ganglioside (NeuGc) monoclonal antibody. G-4 was recognized by the monoclonal ant ibody which reacts with I-active ganglioside (NeuGc). In addition, sia lic acid moiety of the gangliosides from T. sergenti piroplasms was al so analyzed. N-acetylneuraminic acid-containing gangliosides were hard ly 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 moie ty. These results suggest that G-1, G-2, G-3, and G-4 gangliosides are GM3 (NeuGc) [NeuGc alpha 2-3Gal beta 1-4Glc beta 1-1Cer], SPG (NeuGc) [NeuGc alpha 2-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1Ce r], i-active ganglioside (NeuGc) [NeuGc alpha 2-3Gal beta 1-4GlcNAc be ta 1-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1Cer], and I-a ctive ganglioside(NeuGc) [NeuGc alpha 2-3Gal beta 1-4GlcNAc beta 1-3 ( Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-6) Gal beta 1-4GlcNAc beta 1-3G al beta 1-4Glc beta 1-1Cer], respectively.