Synaptic vesicle proteins under conditions of rest and activation: analysis by 2-D difference gel electrophoresis.
Synaptic vesicles are organelles of the nerve terminal that secrete neurotransmitters by fusion with the presynaptic plasma membrane. Vesicle fusion is tightly controlled by depolarization of the plasma membrane and a set of proteins that may undergo post-translational modifications such as phosphorylation. In order to identify proteins that undergo modifications as a result of synaptic activation, we induced massive exocytosis and analysed the synaptic vesicle compartment by benzyldimethyl-n-hexadecylammonium chloride (BAC)/SDS-PAGE and difference gel electrophoresis (DIGE) followed by MALDI-TOF-MS. We identified eight proteins that revealed significant changes in abundance following nerve terminal depolarization. Of these, six were increased and two were decreased in abundance. Three of these proteins were phosphorylated as detected by Western blot analysis. In addition, we identified an unknown synaptic vesicle protein whose abundance increased on synaptic activation. Our results demonstrate that depolarization of the presynaptic compartment induces changes in the abundance of synaptic vesicle proteins and post-translational protein modification.
Burré
Jacqueline
J
Department of Neurochemistry, JW Goethe-University, Frankfurt/Main, Germany. j.burre@zoology.uni-frankfurt.de
Beckhaus
Tobias
T
Corvey
Carsten
C
Karas
Michael
M
Zimmermann
Herbert
H
Volknandt
Walter
W
D000818:Animals; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005089:Exocytosis / Q000187:drug effects; D005233:Fatty Alcohols; D009419:Nerve Tissue Proteins / Q000032:analysis*; D010750:Phosphoproteins / Q000032:analysis; D010766:Phosphorylation; D000644:Quaternary Ammonium Compounds; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013572:Synaptic Vesicles / Q000737:chemistry* / Q000187:drug effects / Q000502:physiology*; D050825:Synaptosomal-Associated Protein 25 / Q000032:analysis; D050988:Vesicle-Associated Membrane Protein 2 / Q000032:analysis
D005233:Fatty Alcohols; D009419:Nerve Tissue Proteins; D010750:Phosphoproteins; D000644:Quaternary Ammonium Compounds; C494210:Snap25 protein, rat; D050825:Synaptosomal-Associated Protein 25; C494706:Vamp2 protein, rat; D050988:Vesicle-Associated Membrane Protein 2; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; C012817:cetalkonium chloride