Gallo J M, Anderton B H
EMBO J. 1983;2(4):479-83. doi: 10.1002/j.1460-2075.1983.tb01450.x.
Two hybridoma cell lines were selected after the fusion of the myeloma cell line X-63 Ag8-653 with spleen cells from mice immunized with bovine brain microtubules. These lines, clones 3F3 and 16D3, secrete IgM antibodies both staining a fibrillar network in fibroblasts. Autoradiography of immunoblots of SDS gels showed that the antigenic determinants defined by these antibodies are present on tubulin and also on several other polypeptides in mammalian cells. In contrast, they were found to react only with tubulin in Trypanosoma brucei, parasitic protozoan which are the causative agent of sleeping sickness. By immunofluorescence microscopy, 3F3 bound only to a subpopulation of microtubules associated with the flagellum of these cells when, under the same conditions, 16D3 stained other microtubule populations including sub-pellicular microtubules. These results show that flagellar tubulin differs from tubulin of other locations in the same cell by at least one antigenic determinant which could be involved in microtubule specialization.
骨髓瘤细胞系X-63 Ag8-653与用牛脑微管免疫的小鼠脾细胞融合后,筛选出了两个杂交瘤细胞系。这两个细胞系,即克隆3F3和16D3,分泌的IgM抗体均能使成纤维细胞中的纤维状网络染色。SDS凝胶免疫印迹的放射自显影显示,这些抗体所定义的抗原决定簇存在于微管蛋白上,也存在于哺乳动物细胞中的其他几种多肽上。相比之下,发现它们仅与布氏锥虫(昏睡病的病原体,一种寄生原生动物)中的微管蛋白发生反应。通过免疫荧光显微镜观察,当在相同条件下16D3能使包括表膜下微管在内的其他微管群体染色时,3F3仅与这些细胞鞭毛相关的一部分微管结合。这些结果表明,鞭毛微管蛋白与同一细胞中其他位置的微管蛋白至少在一个抗原决定簇上存在差异,这一抗原决定簇可能与微管的特化有关。