The proteome of the presynaptic active zone: from docked synaptic vesicles to adhesion molecules and maxi-channels.
The presynaptic proteome controls neurotransmitter release and the short and long term structural and functional dynamics of the nerve terminal. Using a monoclonal antibody against synaptic vesicle protein 2 we immunopurified a presynaptic compartment containing the active zone with synaptic vesicles docked to the presynaptic plasma membrane as well as elements of the presynaptic cytomatrix. Individual protein bands separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were subjected to nanoscale-liquid chromatography electrospray ionization-tandem mass spectrometry. Combining this method with 2-dimensional benzyldimethyl-n-hexadecylammonium chloride/sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization time of flight and immunodetection we identified 240 proteins comprising synaptic vesicle proteins, components of the presynaptic fusion and retrieval machinery, proteins involved in intracellular signal transduction, a large variety of adhesion molecules and proteins potentially involved in regulating the functional and structural dynamics of the pre-synapse. Four maxi-channels, three isoforms of voltage-dependent anion channels and the tweety homolog 1 were co-isolated with the docked synaptic vesicles. As revealed by in situ hybridization, tweety homolog 1 reveals a distinct expression pattern in the rodent brain. Our results add novel information to the proteome of the presynaptic active zone and suggest that in particular proteins potentially involved in the short and long term structural modulation of the mature presynaptic compartment deserve further detailed analysis.
Morciano
Marco
M
Neurochemistry, Biocenter of Goethe-University, Frankfurt am Main, Germany.
Beckhaus
Tobias
T
Karas
Michael
M
Zimmermann
Herbert
H
Volknandt
Walter
W
D000818:Animals; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D007120:Immunochemistry; D007136:Immunoglobulins / Q000276:immunology; D017403:In Situ Hybridization; D007473:Ion Channels / Q000235:genetics*; D008297:Male; D008854:Microscopy, Electron; D033681:Mitochondrial Membrane Transport Proteins; D020543:Proteome / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D017661:Receptors, Presynaptic / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013572:Synaptic Vesicles / Q000235:genetics*; D050995:Voltage-Dependent Anion Channel 1 / Q000235:genetics; D050996:Voltage-Dependent Anion Channel 2 / Q000235:genetics; D050994:Voltage-Dependent Anion Channels / Q000235:genetics
D015815:Cell Adhesion Molecules; D007136:Immunoglobulins; D007473:Ion Channels; D033681:Mitochondrial Membrane Transport Proteins; D020543:Proteome; D017661:Receptors, Presynaptic; C494764:Vdac1 protein, rat; C494767:Vdac2 protein, rat; C494769:Vdac3 protein, rat; D050996:Voltage-Dependent Anion Channel 2; D050994:Voltage-Dependent Anion Channels; D050995:Voltage-Dependent Anion Channel 1