title: Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins.
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.
authors.affiliation: 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.
authors.forename: Benjamin G
authors.initials: BG
authors.lastname: Wilhelm
authors.affiliation: Bioanalytical Mass Spectrometry Group, Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
authors.forename: Sunit
authors.initials: S
authors.lastname: Mandad
authors.affiliation: 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.
authors.forename: Sven
authors.initials: S
authors.lastname: Truckenbrodt
authors.affiliation: 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.
authors.forename: Katharina
authors.initials: K
authors.lastname: Kröhnert
authors.affiliation: 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.
authors.forename: Christina
authors.initials: C
authors.lastname: Schäfer
authors.affiliation: 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.
authors.forename: Burkhard
authors.initials: B
authors.lastname: Rammner
authors.affiliation: Leibniz Institut für Molekulare Pharmakologie, Department of Molecular Pharmacology and Cell Biology, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Seong Joo
authors.initials: SJ
authors.lastname: Koo
authors.affiliation: Leibniz Institut für Molekulare Pharmakologie, Department of Molecular Pharmacology and Cell Biology, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Gala A
authors.initials: GA
authors.lastname: Claßen
authors.affiliation: Leibniz Institut für Molekulare Pharmakologie, Department of Molecular Pharmacology and Cell Biology, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Krauss
authors.affiliation: Leibniz Institut für Molekulare Pharmakologie, Department of Molecular Pharmacology and Cell Biology, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Haucke
authors.affiliation: 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.
authors.forename: Henning
authors.initials: H
authors.lastname: Urlaub
authors.affiliation: 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.
authors.forename: Silvio O
authors.initials: SO
authors.lastname: Rizzoli
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D005089:Exocytosis; D021621:Imaging, Three-Dimensional; D015151:Immunoblotting / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods; D008959:Models, Neurological; D017729:Presynaptic Terminals / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D021381:Protein Transport; D051381:Rats; D017208:Rats, Wistar; D013572:Synaptic Vesicles / Q000737:chemistry / Q000378:metabolism*; D013574:Synaptosomes / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D033921:Vesicular Transport Proteins / Q000032:analysis / Q000378:metabolism*
chemical_list: D033921:Vesicular Transport Proteins