Dragon E A, Brothers V M, Wrightsman R A, Manning J
Mol Biochem Parasitol. 1985 Sep;16(3):213-29. doi: 10.1016/0166-6851(85)90065-9.
Trypanosoma cruzi (Peru strain) trypomastigotes and epimastigotes were biosynthetically labeled with [35S]methionine, and the proteins were analyzed by two dimensional polyacrylamide gel electrophoresis (2D-PAGE). 2D-PAGE analysis of the trypomastigotes showed a complex array of polypeptides with distinct clusters at Mr 88 000-92 000, isoelectric point (pI) 5.6-6.0, and Mr 72 000-76 000, pI 5.6-5.8. 2D-PAGE analysis of the epimastigotes did not show the cluster of polypeptides at Mr 90 000. When the trypomastigote lysate was reacted with sera from either mice or humans chronically infected with T. cruzi, 10-50 polypeptides were immunoprecipitated. Five of these polypeptides were recognized by all sera tested. However, of these polypeptides, only three, two of Mr 90 000 and one of Mr 150 000, can be identified by immunoreaction of [35S]methionine-labeled live parasites as surface proteins of T. cruzi trypomastigotes. 125I-iminobiotinylated surface proteins isolated from T. cruzi trypomastigotes were immunoprecipitated with the same series of sera as described above. Chagasic sera immunoprecipitated an antigen of Mr 90 000. The [35S]methionine and 125I-labeled Mr 90 000 polypeptides were not immunoprecipitated with sera from individuals infected with Leishmania donovani, Leishmania braziliensis, Leishmania tropica or Leishmania mexicana. These data indicate that a surface polypeptide of Mr 90000, pI 5.8-5.9 is a viable candidate for a Chagas' disease diagnostic antigen.
克氏锥虫(秘鲁株)的锥鞭毛体和上鞭毛体用[35S]甲硫氨酸进行生物合成标记,蛋白质通过二维聚丙烯酰胺凝胶电泳(2D-PAGE)进行分析。锥鞭毛体的2D-PAGE分析显示出一系列复杂的多肽,在分子量(Mr)88000 - 92000、等电点(pI)5.6 - 6.0以及Mr 72000 - 76000、pI 5.6 - 5.8处有明显的簇。上鞭毛体的2D-PAGE分析未显示出Mr 90000处的多肽簇。当锥鞭毛体裂解物与慢性感染克氏锥虫的小鼠或人类血清反应时,有10 - 50种多肽被免疫沉淀。所有测试血清都能识别其中5种多肽。然而,在这些多肽中,只有3种,即两种Mr 90000的和一种Mr 150000的,可通过[35S]甲硫氨酸标记的活寄生虫的免疫反应鉴定为克氏锥虫锥鞭毛体的表面蛋白。从克氏锥虫锥鞭毛体分离的125I - 亚氨基生物素化表面蛋白用上述相同系列的血清进行免疫沉淀。恰加斯病血清免疫沉淀出一种Mr 90000的抗原。[35S]甲硫氨酸和125I标记的Mr 90000多肽不能被感染杜氏利什曼原虫、巴西利什曼原虫、热带利什曼原虫或墨西哥利什曼原虫的个体的血清免疫沉淀。这些数据表明,一种Mr 90000、pI 5.8 - 5.9的表面多肽是恰加斯病诊断抗原的一个可行候选物。