Abuin G, Couto A S, de Lederkremer R M, Casal O L, Galli C, Colli W, Alves M J
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Brazil.
Exp Parasitol. 1996 Apr;82(3):290-7. doi: 10.1006/expr.1996.0036.
Tc-85, an 85-kDa surface glycoprotein specific for the trypomastigote stage of Trypanosoma cruzi, has been implicated in the invasion of host cells by the parasite. Radioactive palmitic acid was incorporated into Tc-85 immunoprecipitated from the culture medium with the H1A10 monoclonal antibody, suggesting that shedding occurs with Tc-85 bearing its GPI anchor. In contrast to the glycoprotein remaining in the parasites, the glycosylphosphatidylinositol moiety in shed Tc-85 is resistant to phosphatidylinositol phospholipase C and becomes susceptible to the enzyme following alkali treatment. An alkylglycerol was identified by thin layer chromatography of an ether extract after the enzymatic reaction. Resistance to cleavage by phospholipase C is due to fatty acid esterification of the inositol residue in shed Tc-85. This is the first example of inositol modification in anchors from a glycoprotein of Trypanosoma cruzi.
Tc-85是一种分子量为85 kDa的表面糖蛋白,对克氏锥虫的锥鞭毛体阶段具有特异性,它与该寄生虫对宿主细胞的入侵有关。用H1A10单克隆抗体从培养基中免疫沉淀得到的Tc-85掺入了放射性棕榈酸,这表明Tc-85在携带其糖基磷脂酰肌醇(GPI)锚时会发生脱落。与留在寄生虫体内的糖蛋白不同,脱落的Tc-85中的糖基磷脂酰肌醇部分对磷脂酰肌醇磷脂酶C具有抗性,而在碱处理后会变得对该酶敏感。酶促反应后,通过醚提取物的薄层色谱法鉴定出一种烷基甘油。对磷脂酶C切割的抗性是由于脱落的Tc-85中肌醇残基的脂肪酸酯化。这是克氏锥虫糖蛋白锚中肌醇修饰的首个例子。