Foisner R, Wiche G
J Ultrastruct Res. 1985 Oct-Nov;93(1-2):33-41. doi: 10.1016/0889-1605(85)90083-7.
The effects of taxol on microtubule-associated proteins of high molecular weight (MAPs) were studied in vitro. After negative staining, microtubules reconstituted in the presence of taxol from preparations of partially purified tubulin and MAPs, besides being bundled, displayed prominent elongated or globular extensions without apparent regularity. These extensions, but not the tubulin polymer, were heavily decorated after immuno-gold-labeling using antibodies to MAP-1 and MAP-2. Microtubules reconsituted in the absence of taxol showed a much more regular, and apparently helical, arrangement of MAPs along their surfaces. The formation of polymeric structures was also observed when preparation of MAPs free of tubulin were incubated with taxol. In this case in addition to large network-type aggregates with little apparent substructure, more regular structures seemingly consisting of approximately 5-nm-thick filaments arrayed in parallel were observed. Taxol-induced MAP aggregation occurred rapidly and was directly proportional to the concentration of protein, as revealed by optical density measurements. It is concluded that taxol, aside from promoting the assembly of tubulin and stabilizing microtubules, promotes MAP/MAP interaction.
在体外研究了紫杉醇对高分子量微管相关蛋白(MAPs)的影响。负染后,在紫杉醇存在的情况下,由部分纯化的微管蛋白和MAPs制剂重构的微管,除了成束外,还显示出突出的、无明显规则的细长或球状延伸。使用针对MAP-1和MAP-2的抗体进行免疫金标记后,这些延伸部分而非微管蛋白聚合物被大量标记。在没有紫杉醇的情况下重构的微管显示出MAPs沿着其表面更规则且明显呈螺旋状的排列。当不含微管蛋白的MAPs制剂与紫杉醇一起孵育时,也观察到了聚合物结构的形成。在这种情况下,除了几乎没有明显亚结构的大型网络型聚集体外,还观察到了似乎由排列成平行的约5纳米厚细丝组成的更规则结构。如光密度测量所示,紫杉醇诱导的MAP聚集迅速发生,且与蛋白质浓度成正比。得出的结论是,紫杉醇除了促进微管蛋白组装和稳定微管外,还促进MAP/MAP相互作用。