Trinczek B, Marx A, Mandelkow E M, Murphy D B, Mandelkow E
Max-Planck-Unit for Structural Molecular Biology, Hamburg, Germany.
Mol Biol Cell. 1993 Mar;4(3):323-35. doi: 10.1091/mbc.4.3.323.
Microtubules can adjust their length by the mechanism of dynamic instability, that is by switching between phases of growth and shrinkage. Thus far this phenomenon has been studied with microtubules that contain several components, that is, a mixture of tubulin isoforms, with or without a mixture of microtubule-associated proteins (MAPs), which can act as regulators of dynamic instability. Here we concentrate on the influence of the tubulin component. We have studied MAP-free microtubules from the marginal band of avian erythrocytes and compared them with mammalian brain microtubules. The erythrocyte system was selected because it represents a naturally stable aggregate of microtubules; second, the tubulin is largely homogeneous, in contrast to brain tubulin. Qualitatively, erythrocyte microtubules show similar features as brain microtubules, but they were found to be much less dynamic. The critical concentration of elongation, and the rates of association and dissociation of tubulin are all lower than with brain microtubules. Catastrophes are rare, rescues frequent, and shrinkage slow. This means that dynamic instability can be controlled by the tubulin isotype, independently of MAPs. Moreover, the extent of dynamic behavior is highly dependent on buffer conditions. In particular, dynamic instability is strongly enhanced in phosphate buffer, both for erythrocyte marginal band and brain microtubules. The lower stability in phosphate buffer argues against the hypothesis that a cap of tubulin.GDP.Pi subunits stabilizes microtubules. The difference in dynamics between tubulin isotypes and between the two ends of microtubules is preserved in the different buffer systems.
微管可以通过动态不稳定性机制来调节其长度,即通过在生长和收缩阶段之间切换。到目前为止,这种现象已经在包含多种成分的微管中进行了研究,也就是说,是在含有或不含有微管相关蛋白(MAPs)混合物的微管蛋白亚型混合物中进行研究的,微管相关蛋白可以作为动态不稳定性的调节因子。在这里,我们专注于微管蛋白成分的影响。我们研究了来自鸟类红细胞边缘带的无MAPs微管,并将它们与哺乳动物脑微管进行了比较。选择红细胞系统是因为它代表了微管的天然稳定聚集体;其次,与脑微管蛋白相比,微管蛋白在很大程度上是同质的。定性地说,红细胞微管显示出与脑微管相似的特征,但发现它们的动态性要低得多。微管蛋白的伸长临界浓度、结合和解离速率均低于脑微管。灾难性事件很少发生,救援频繁,收缩缓慢。这意味着动态不稳定性可以由微管蛋白同种型控制,而与MAPs无关。此外,动态行为的程度高度依赖于缓冲条件。特别是,对于红细胞边缘带和脑微管,在磷酸盐缓冲液中动态不稳定性都会显著增强。磷酸盐缓冲液中较低的稳定性与微管蛋白.GDP.Pi亚基帽稳定微管的假设相悖。微管蛋白同种型之间以及微管两端之间的动力学差异在不同的缓冲系统中得以保留。