Wiche G, Cole R D
Proc Natl Acad Sci U S A. 1976 Apr;73(4):1227-31. doi: 10.1073/pnas.73.4.1227.
Tubulin from cultures of the rat glial cell clone C6 could be polymerized in vitro into intact microtubules. The polymerization was reversible and spontaneous, i.e., no addition of heterologous nucleation centers was necessary. Two cycles of polymerization/depolymerization yielded tubulin preparations of 95% purity as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Electron microscopy was used to show that the microtubules assembled in vitro by two cycles of polymerization/depolymerization were morphologically intact and temperature sensitive. In contrast, tubulin from neuroblastoma cells, clone Neuro-2A, could not be polymerized in a reversible fashion. The discovery of a cell line from which tubulin can be reversibly polymerized in vitro establishes a model system for studies of cell-cycle- and cell-type-dependent regulatory mechanisms controlling the assembly of microtubules.
大鼠神经胶质细胞克隆C6培养物中的微管蛋白能够在体外聚合成完整的微管。这种聚合是可逆且自发的，也就是说，无需添加异源成核中心。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定，经过两个聚合/解聚循环后得到的微管蛋白制剂纯度达95%。电子显微镜显示，通过两个聚合/解聚循环在体外组装的微管在形态上是完整的且对温度敏感。相比之下，神经母细胞瘤细胞克隆Neuro-2A中的微管蛋白无法以可逆方式聚合。发现一种其微管蛋白可在体外进行可逆聚合的细胞系，为研究控制微管组装的细胞周期和细胞类型依赖性调节机制建立了一个模型系统。
