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Isolation of intraflagellar transport trains

机译：鞭毛内运输火车的隔离

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The intraflagellar transport (IFT) system was first identified in situ by electron microscopy in thin sections of plastic-embedded flagella as linear arrays of electrondense particles, located between the B tubules of the outer doublets and the flagellar membrane. These arrays of particles are referred to as IFT trains. Upon membrane rupture, IFT trains are thought to easily dissociate to yield soluble IFT particles, which are commonly purified through sucrose gradients as ~16-17S complexes. The latters easily dissociate into two subcomplexes, named A and B. We report here the isolation, visualization, and identification by immunolabeling of flexible strings of IFT particles, which are structurally similar to in situ IFT trains and appear to be formed by both complex A and complex B polypeptides. Moreover, the particles forming isolated IFT trains are structurally similar to the individual particles found in the ~17S gradient peak. Our results provide the first direct evidence that ~17S particles do indeed compose the IFT trains. The paper also represents the first isolation of the IFT trains, and opens new possibilities for higher resolution studies on their structure and how particles are attached to each other to form the particle trains.
机译：鞭毛内转运（IFT）系统首先通过电子显微镜在塑料包埋的鞭毛的薄切片中原位鉴定为位于双合体B小管和鞭毛膜之间的电子致密颗粒的线性阵列。这些粒子阵列称为IFT列。膜破裂后，人们认为IFT序列很容易解离，产生可溶的IFT颗粒，通常通过蔗糖梯度将其纯化为约16-17S的复合物。后者容易分解为两个亚复合物，分别称为A和B。我们在这里报告了通过免疫标记IFT颗粒的细线进行的分离，可视化和鉴定，这些结构与原位IFT序列相似，并且似乎由两种复合物A形成和复杂的B多肽。此外，形成隔离的IFT列的粒子在结构上类似于在〜17S梯度峰中发现的单个粒子。我们的结果提供了第一个直接证据，表明〜17S颗粒确实构成了IFT序列。该论文还代表了IFT列的第一个隔离，并为高分辨率研究IFT列的结构以及如何将粒子彼此附着以形成粒子列开辟了新的可能性。

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