Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins

被引:524
|
作者
Wilhelm, Benjamin G. [1 ,2 ]
Mandad, Sunit [3 ]
Truckenbrodt, Sven [1 ,5 ]
Kroehnert, Katharina [1 ]
Schaefer, Christina [1 ]
Rammner, Burkhard [1 ]
Koo, Seong Joo [6 ]
Classen, Gala A. [6 ]
Krauss, Michael [6 ]
Haucke, Volker [6 ]
Urlaub, Henning [3 ,4 ]
Rizzoli, Silvio O. [1 ]
机构
[1] Univ Gottingen, European Neurosci Inst, Cluster Excellence Nanoscale Microscopy & Mol Phy, Med Ctr,Dept Neuro & Sensory Physiol, D-37073 Gottingen, Germany
[2] Int Max Planck Res Sch Neurosci, D-37077 Gottingen, Germany
[3] Max Planck Inst Biophys Chem, Bioanalyt Mass Spectrometry Grp, D-37077 Gottingen, Germany
[4] Univ Med Ctr Gottingen, Dept Clin Chem, D-37075 Gottingen, Germany
[5] Int Max Planck Res Sch Mol Biol, D-37077 Gottingen, Germany
[6] Leibniz Inst Mol Pharmacol, Dept Mol Pharmacol & Cell Biol, D-13125 Berlin, Germany
基金
欧洲研究理事会;
关键词
NEUROTRANSMITTER RELEASE; SYNTAXIN CLUSTERS; MEMBRANE; FUSION; POOL; QUANTIFICATION; EXOCYTOSIS; SYNAPSES; ENDOCYTOSIS; SUFFICIENT;
D O I
10.1126/science.1252884
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页码:1023 / 1028
页数:6
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