Synapsin II. Mapping of a domain in the NH2-terminal region which binds to small synaptic vesicles.
The synapsins are a family of neuron-specific phosphoproteins that selectively bind to small synaptic vesicles in the presynaptic nerve terminal. Using the cDNA encoding rat synapsin IIb, we employed an Escherichia coli expression system to synthesize a variety of fusion proteins containing a truncated protein A linked to different portions of the NH2-terminal region of synapsin II. The recombinant proteins were purified by IgG-Sepharose chromatography and tested in vitro for their ability to bind to purified synaptic vesicles. These experiments identified a region between amino acids 43 and 121 of the amino-terminal portion of synapsin II which binds to synaptic vesicles. Mild trypsinization of synaptic vesicles reduces binding of recombinant proteins to synaptic vesicles, suggesting that the interaction between synapsin II and the vesicles is in part mediated by a synaptic vesicle protein. The 42 NH2-terminal amino acids of synapsin II are not necessary for binding to synaptic vesicles, although this domain contains the phosphorylation site for cAMP-dependent protein kinase.
Thiel
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G
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021.
Südhof
T C
TC
Greengard
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P
D000818:Animals; D002540:Cerebral Cortex / Q000378:metabolism; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D009479:Neuropeptides / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000235:genetics*; D010766:Phosphorylation; D010957:Plasmids; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D016704:Synapsins; D013572:Synaptic Vesicles / Q000378:metabolism*
D009479:Neuropeptides; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D016704:Synapsins