Couto A S, Katzin A M, Colli W, de Lederkremer R M
Departamento de Bioquímica, Universidade de São Paulo, Brazil.
Mol Biochem Parasitol. 1987 Nov;26(1-2):145-53. doi: 10.1016/0166-6851(87)90138-1.
The carbohydrate moiety of the Tc-85 surface glycoprotein from the infective trypomastigote form of Trypanosoma cruzi was analysed. Tc-85 could be metabolically labeled by incubation of the cells with D-[14C]mannose or D-[14C]glucose. Degradation techniques were performed directly on the polyacrylamide gel band containing labeled Tc-85. A mannobiose was cleaved by beta-elimination and further treatment of the remaining material under conditions which liberate N-asparaginyl linkages, released a complex oligosaccharide. The presence of sialic acid was demonstrated by: mild acid hydrolysis, neuraminidase treatment and periodate oxidation under mild conditions followed by NaB3H4 reduction, hydrolysis, and detection of NANA7 by paper electrophoresis. In addition, the chromatographic behavior of the asialooligosaccharide was significantly different from that of the original sample. Galactose, mannose and glucosamine are the other monosaccharide components of the sialooligosaccharide.
对克氏锥虫感染性锥鞭毛体形式的Tc - 85表面糖蛋白的碳水化合物部分进行了分析。通过将细胞与D - [¹⁴C]甘露糖或D - [¹⁴C]葡萄糖一起孵育,Tc - 85可被代谢标记。降解技术直接在含有标记Tc - 85的聚丙烯酰胺凝胶条带上进行。通过β - 消除裂解出一个甘露二糖,并且在释放N - 天冬酰胺键的条件下对剩余物质进行进一步处理,释放出一种复合寡糖。通过以下方法证实了唾液酸的存在:温和酸水解、神经氨酸酶处理以及在温和条件下进行高碘酸盐氧化,随后用NaB₃H₄还原、水解,并通过纸电泳检测NANA7。此外,去唾液酸寡糖的色谱行为与原始样品有显著差异。半乳糖、甘露糖和氨基葡萄糖是去唾液酸寡糖的其他单糖成分。