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PMID: 6404912 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Synapsin I (protein I), a nerve terminal-specific phosphoprotein. III. Its association with synaptic vesicles studied in a highly purified synaptic vesicle preparation.

The Journal of cell biology ·Vol. 96 ·No. 5 ·1983-05-00 ·Pages 1374-88

Huttner WB, Schiebler W, Greengard P, De Camilli P

Abstract

Synapsin I (protein I) is a neuron-specific phosphoprotein, which is a substrate for cAMP-dependent and Ca/calmodulin-dependent protein kinases. In two accompanying studies (De Camilli, P., R. Cameron, and P. Greengard, and De Camilli, P., S. M. Harris, Jr., W. B. Huttner, and P. Greengard, 1983, J. Cell Biol. 96:1337-1354 and 1355-1373) we have shown, by immunocytochemical techniques at the light microscopic and electron microscopic levels, that synapsin I is present in the majority of, and possibly in all, nerve terminals, where it is primarily associated with synaptic vesicles. In the present study we have prepared a highly purified synaptic vesicle fraction from rat brain by a procedure that involves permeation chromatography on controlled-pore glass as a final purification step. Using immunological methods, synapsin I concentrations were determined in various subcellular fractions obtained in the course of vesicle purification. Synapsin I was found to copurify with synaptic vesicles and to represent approximately 6% of the total protein in the highly purified synaptic vesicle fraction. The copurification of synapsin I with synaptic vesicles was dependent on the use of low ionic strength media throughout the purification. Synapsin I was released into the soluble phase by increased ionic strength at neutral pH, but not by nonionic detergents. The highly purified synaptic vesicle fraction contained a calcium-dependent protein kinase that phosphorylated endogenous synapsin I in its collagenase-sensitive tail region. The phosphorylation of this region appeared to facilitate the dissociation of synapsin I from synaptic vesicles under the experimental conditions used.

MeSH Terms
Animals Cell Fractionation Cerebral Cortex/analysis,ultrastructure Clathrin Membrane Proteins/analysis Microscopy, Electron Nerve Tissue Proteins/analysis Phosphoproteins/analysis Phosphorylation Rats Rats, Inbred Strains Solubility Synapsins Synaptic Vesicles/analysis Tissue Distribution
Chemicals
Clathrin Membrane Proteins Nerve Tissue Proteins Phosphoproteins Synapsins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Huttner W B
Schiebler W
Greengard P
De Camilli P
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35 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.96.5.1374
Published
1983-05-00
Pages
1374-88
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2112660
Subset
IM
Grants
NIMH NIH HHS · MH-17387 · United States
NINDS NIH HHS · NS-08440 · United States
External Links
PubMed source DOI: 10.1083/jcb.96.5.1374 PMC Full Text View journal details
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