Singh B N
Department of Microbiology and Immunology, SUNY Health Science Center, Syracuse 13210.
Mol Biochem Parasitol. 1993 Feb;57(2):281-94. doi: 10.1016/0166-6851(93)90204-b.
Lipophosphoglycan-like glycoconjugates were isolated, purified and partially characterized from Tritrichomonas foetus and Trichomonas vaginalis. Cell surface radiolabeling of both trichomonads by the galactose oxidase/NaB[3H]4 technique indicated that the glycoconjugate was located on the cell surface of the parasites. The glycoconjugates were extracted from the delipidated residue fraction with the solvent, water/ethanol/diethylether/pyridine/NH4OH (15:15:5:1:0.017) and were purified to homogeneity by Sepharose CL-4B followed by octyl-Sepharose chromatography and methanol precipitation. The glycoconjugates migrated as broad bands upon SDS-PAGE. The T. foetus glycoconjugate contained large amounts of fucose along with some mannose, galactose, glucosamine and glucose and trace amounts of galactosamine and inositol. The T. vaginalis glycoconjugate appeared to contain large amounts of glucosamine and galactose along with some glucose, mannose and traces of galactosamine and inositol. The surface-labeled glycoconjugates from both parasites was found to be deaminated with nitrous acid and susceptible to phosphatidylinositol-specific phospholipase C, indicating the presence of a phospholipid anchor. Furthermore, these glycoconjugate were found to contain phosphate and were labile to hydrolysis by mild acid, strongly suggesting that the intact molecule is related to Leishmania lipophosphoglycans (LPG). The most striking and the unique features of these glycoconjugate molecules are the presence of large amounts of fucose in T. foetus and glucosamine in T. vaginalis along with the presence of galactosamine in both parasites. These results indicate that these glycoconjugates are new types of LPG-like molecules expressed on the trichomonad cell surface and are structurally distinct from Leishmania LPG.
从胎儿三毛滴虫和阴道毛滴虫中分离、纯化并对脂磷壁酸样糖缀合物进行了部分特性鉴定。通过半乳糖氧化酶/NaB[3H]4技术对两种滴虫进行细胞表面放射性标记,结果表明该糖缀合物位于寄生虫的细胞表面。用溶剂水/乙醇/二乙醚/吡啶/NH4OH(15:15:5:1:0.017)从脱脂残渣部分提取糖缀合物,并通过琼脂糖CL-4B,随后进行辛基琼脂糖层析和甲醇沉淀将其纯化至同质。该糖缀合物在SDS-PAGE上迁移为宽带。胎儿三毛滴虫糖缀合物含有大量岩藻糖,以及一些甘露糖、半乳糖、氨基葡萄糖和葡萄糖,还有痕量的半乳糖胺和肌醇。阴道毛滴虫糖缀合物似乎含有大量氨基葡萄糖和半乳糖,以及一些葡萄糖、甘露糖和痕量的半乳糖胺和肌醇。发现两种寄生虫表面标记的糖缀合物都能用亚硝酸脱氨,并且对磷脂酰肌醇特异性磷脂酶C敏感,表明存在磷脂锚。此外,这些糖缀合物被发现含有磷酸盐,并且对弱酸水解不稳定,这强烈表明完整分子与利什曼原虫脂磷壁酸(LPG)有关。这些糖缀合物分子最显著和独特的特征是胎儿三毛滴虫中存在大量岩藻糖,阴道毛滴虫中存在氨基葡萄糖,以及两种寄生虫中都存在半乳糖胺。这些结果表明,这些糖缀合物是在滴虫细胞表面表达的新型LPG样分子,在结构上与利什曼原虫LPG不同。