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Science. 2014 May 30;344(6187):1023-8. doi: 10.1126/science.1252884.

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

Author information

1
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.
2
Bioanalytical Mass Spectrometry Group, Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
3
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 Molecular Biology, 37077 Göttingen, Germany.
4
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.
5
Leibniz Institut für Molekulare Pharmakologie, Department of Molecular Pharmacology and Cell Biology, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
6
Bioanalytical Mass Spectrometry Group, Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen, Germany. Bioanalytics, Department of Clinical Chemistry, University Medical Center Göttingen, 37075 Göttingen, Germany.
7
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. srizzol@gwdg.de.

Abstract

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.

Comment in

PMID:
24876496
DOI:
10.1126/science.1252884
[Indexed for MEDLINE]
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