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首页> 外文期刊>Journal of Molecular Biology >Visualization of the stop of microtubule depolymerization that occurs at the high-density region of microtubule-associated protein 2 (MAP2)
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Visualization of the stop of microtubule depolymerization that occurs at the high-density region of microtubule-associated protein 2 (MAP2)

机译：可视化发生在微管相关蛋白2（MAP2）的高密度区域的微管解聚停止

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Individual microtubules (MTs) repeat alternating phases of polymerization and depolymerization, a process known as dynamic instability. Microtubule-associated proteins (MAPS) regulate the dynamic instability by increasing the rescue frequency. To explore the influence of MAP2 on in vitro MT dynamics, we correlated the distribution of MAP2 on individual MTs with the dynamic phase changes of the same MTs,MAP2 was modified selectively on its projection region by X-rhodamine iodoacetamide without altering the MT-binding activity. When the labeled MAP2 was added to MTs, the fluorescence was distributed along almost the entire length of individual MTs. However, the inhomogeneity of the distribution gradually became obvious due to the fluorescence bleaching, and the MTs appeared to consist of rapidly bleached portions (RBPs) and slowly bleached portions (SBPs), which were distributed randomly along the MT. By measuring the duration of fluorescence bleaching, the density of MAP2 in SBP was estimated to be approximately 2.5 times higher than the RBP. The average tubulin:MAP2 ratio in SBP was calculated to be 16. When the MT dynamics were observed by dark-field microscopy after determining the MAP2 distribution, rescues were always found to occur only at the SBPs. Ws also displayed intermittent shortening by repeated depolymerization phases separated by pause phases. In these cases, depolymerization phases stopped only at the SBPs. Not every SBP stopped depolymerization, but depolymerization always stopped at an SBP. Taken together, we suggest that there is a minimum density of MAP2 that is necessary to stop depolymerization.
机译：各个微管（MT）重复进行聚合和解聚的交替阶段，这一过程称为动态不稳定性。微管相关蛋白（MAPS）通过增加救援频率来调节动态不稳定性。为了探讨MAP2对体外MT动力学的影响，我们将MAP2在单个MT上的分布与相同MT的动态相变相关联，X-若丹明碘乙酰胺选择性地在其投影区域修饰MAP2，而不会改变MT的结合活动。当将标记的MAP2添加到MT时，荧光几乎沿着各个MT的整个长度分布。然而，由于荧光褪色，分布的不均匀性逐渐变得明显，MT似乎由沿着MT随机分布的快速漂白部分（RBP）和缓慢漂白部分（SBP）组成。通过测量荧光漂白的持续时间，估计SBP中MAP2的密度约为RBP的2.5倍。计算得出SBP中微管蛋白：MAP2的平均比例为16。当在确定MAP2分布后通过暗视野显微镜观察MT动力学时，总是发现仅在SBP发生了抢救。 Ws还显示出间歇性缩短，这是由于重复的解聚相被暂停相隔开。在这些情况下，解聚阶段仅在SBP处停止。并非每个SBP都停止解聚，但是解聚始终在SBP处停止。两者合计，我们建议必须有最小密度的MAP2才能阻止解聚。

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