Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins
Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins
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DOI：
10.1126/science.1252884
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发表时间：
2014-05-30
期刊：
影响因子：
56.9
通讯作者：
Rizzoli, Silvio O.
中科院分区：
文献类型：
--
作者：
Wilhelm, Benjamin G.;Mandad, Sunit;Rizzoli, Silvio O.
长期以来，突触小泡回收一直是细胞运输的一般机制的模型。我们使用了一种综合的方法，结合定量免疫印迹和质谱学来确定蛋白质的数量；电子显微镜来测量细胞器的数量、大小和位置；以及超分辨率荧光显微镜来定位蛋白质。利用这些数据，我们生成了一个“平均”突触的三维模型，显示了300,000个蛋白质的原子细节。突触小泡回收的同一步骤所涉及的蛋白质拷贝数密切相关。相比之下，不同步骤之间的拷贝数变化超过三个数量级，从大约150个拷贝的内体融合蛋白到超过2万个拷贝的胞吐融合蛋白。
Synaptic vesicle recycling has long served as a model for the general mechanisms of cellular trafficking. We used an integrative approach, combining quantitative immunoblotting and mass spectrometry to determine protein numbers; electron microscopy to measure organelle numbers, sizes, and positions; and super-resolution fluorescence microscopy to localize the proteins. Using these data, we generated a three-dimensional model of an "average" synapse, displaying 300,000 proteins in atomic detail. The copy numbers of proteins involved in the same step of synaptic vesicle recycling correlated closely. In contrast, copy numbers varied over more than three orders of magnitude between steps, from about 150 copies for the endosomal fusion proteins to more than 20,000 for the exocytotic ones.