Department of Neuro- and Sensory Physiology, University of Göttingen Medical Center, European Neuroscience Institute, Cluster of Excellence Nanoscale Microscopy and Molecular Physiology of the Brain, Göttingen, Germany. International Max Planck Research School Neurosciences, 37077 Göttingen, Germany.
Bioanalytical Mass Spectrometry Group, Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
Science. 2014 May 30;344(6187):1023-8. doi: 10.1126/science.1252884.
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.
突触囊泡循环一直是细胞运输一般机制的模型。我们采用了一种综合的方法，结合定量免疫印迹和质谱法来确定蛋白质数量；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，以原子细节显示了 30 万个蛋白质。参与突触囊泡再循环同一步骤的蛋白质的拷贝数密切相关。相比之下，不同步骤之间的拷贝数差异超过三个数量级，从大约 150 个内体融合蛋白到超过 20000 个胞吐蛋白。
