Hiller G, Weber K
Cell. 1978 Aug;14(4):795-804. doi: 10.1016/0092-8674(78)90335-5.
A quantitative estimate of the cellular tubulin concentration can be obtained by the use of a radioimmunoassay based upon the competition between tubulin in cell extracts and a known amount of radioactively labeled homogeneous tubulin during binding to a limited amount of anti-tubulin antibodies. This assay shows that a variety of widely used tissue culture cells (mouse L cells, mouse 3T3 cells, chick embryo fibroblasts) have a tubulin content which corresponds to approximately 2.5--3.3% of their total protein. Transformation of mouse 3T3 cells by the DNA virus SV40, and of chick embryo cells by the RNA Rous sarcoma virus, does not change the intracellular tubulin concentration. Transformed cells of brain origin, such as some glia tumor cell lines and some neuroblastoma cell lines, have a much lower tubulin content than does normal brain tissue. The intracellular concentration of tubulin in mouse 3T3 cells is discussed in relation to the number of microtubules detected during interphase by immunofluorescence microscopy. These results are also discussed in view of a mechanism of microtubule elongation in vivo driven by self-assembly.
通过使用一种放射免疫测定法,可以对细胞微管蛋白浓度进行定量估计。该方法基于细胞提取物中的微管蛋白与已知量的放射性标记的同源微管蛋白在与有限量的抗微管蛋白抗体结合过程中的竞争。该测定表明,多种广泛使用的组织培养细胞(小鼠L细胞、小鼠3T3细胞、鸡胚成纤维细胞)的微管蛋白含量约占其总蛋白的2.5% - 3.3%。DNA病毒SV40对小鼠3T3细胞的转化以及RNA劳氏肉瘤病毒对鸡胚细胞的转化,均未改变细胞内微管蛋白浓度。源自脑的转化细胞,如一些胶质细胞瘤细胞系和一些神经母细胞瘤细胞系,其微管蛋白含量比正常脑组织低得多。结合通过免疫荧光显微镜在间期检测到的微管数量,讨论了小鼠3T3细胞中微管蛋白的细胞内浓度。还鉴于体内由自我组装驱动的微管伸长机制对这些结果进行了讨论。