Motile cilia are ancient, evolutionarily conserved organelles whose dysfunction underlies motile ciliopathies, a broad class of human diseases. Motile cilia contain a myriad of different proteins that assemble into an array of distinct machines, and understanding the interactions and functional hierarchies among them presents an important challenge. Here, we defined the protein interactome of motile axonemes using cross-linking mass spectrometry in Tetrahymena thermophila. From over 19,000 cross-links, we identified over 4,700 unique amino acid interactions among over 1,100 distinct proteins, providing both macromolecular and atomic-scale insights into diverse ciliary machines, including the intraflagellar transport system, axonemal dynein arms, radial spokes, the 96-nm ruler, and microtubule inner proteins. Guided by this dataset, we used vertebrate multiciliated cells to reveal functional interactions among several poorly defined human ciliopathy proteins. This dataset provides a resource for studying the biology of an ancient organelle and the molecular etiology of human genetic disease.

中文翻译：

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运动纤毛的氨基酸分辨相互作用组可识别纤毛病蛋白复合物的结构和功能


运动纤毛是古老的、进化上保守的细胞器，其功能障碍是运动纤毛病的基础，这是一大类人类疾病。运动纤毛包含无数不同的蛋白质，这些蛋白质组装成一系列不同的机器，了解它们之间的相互作用和功能层次结构是一项重要的挑战。在这里，我们使用交联质谱法定义了嗜热四膜虫的运动轴丝的蛋白质相互作用组。从 19,000 多个交联中，我们在 1,100 多种不同蛋白质中鉴定了 4,700 多种独特的氨基酸相互作用，为不同的纤毛机器提供了大分子和原子尺度的见解，包括鞭毛内运输系统、轴丝动力蛋白臂、径向辐条、96 nm 标尺和微管内部蛋白。在该数据集的指导下，我们使用脊椎动物多纤毛细胞来揭示几种定义不明确的人类纤毛病蛋白之间的功能相互作用。该数据集为研究古代细胞器的生物学和人类遗传病的分子病因提供了资源。