| Title: | Domain structure of synaptotagmin (p65) |
| Author(s): | Perin MS; Brose N; Jahn R; Sudhof TC; |
| Address: | "Howard Hughes Medical Institute, Dallas, Texas" |
| ISSN/ISBN: | 0021-9258 (Print) 0021-9258 (Linking) |
| Abstract: | "Synaptotagmin (p65) is an abundant and evolutionarily conserved protein of synaptic vesicles that contains two copies of an internal repeat homologous to the regulatory region of protein kinase C. In the current study, we have investigated the biochemical properties of synaptotagmin, demonstrating that it contains five protein domains: an intravesicular amino-terminal domain that is glycosylated but lacks a cleavable signal sequence; a single transmembrane region; a sequence separating the transmembrane region from the two repeats homologous to protein kinase C; the two protein kinase C-homologous repeats; and a conserved carboxyl-terminal sequence following the two repeats homologous to protein kinase C. Sucrose density gradient centrifugations and gel electrophoresis indicate that synaptotagmin monomers associate into dimers and are part of a larger molecular weight complex. A sequence predicted to form an amphipathic alpha-helix that may cause the stable dimerization of synaptotagmin is found in its third domain between the transmembrane region and the protein kinase C-homologous repeats. Synaptotagmin contains a single hypersensitive proteolytic site that is located immediately amino-terminal to the amphipathic alpha-helix, suggesting that synaptotagmin contains a particularly exposed region as the peptide backbone emerges from the dimer. Finally, subcellular fractionation and antibody bead purification demonstrate that synaptotagmin co-purifies with synaptophysin and other synaptic vesicle markers in brain. However, in the adrenal medulla, synaptotagmin was found in both synaptophysin-containing microvesicles and in chromaffin granules that are devoid of synaptophysin, suggesting a shared role for synaptotagmin in the exocytosis of small synaptic vesicles and large dense core catecholaminergic vesicles" |
| Keywords: | Adrenal Medulla/metabolism Amino Acid Sequence Animals Antibodies Antigen-Antibody Complex/analysis Brain/metabolism *Calcium-Binding Proteins Cattle Mating Factor Membrane Glycoproteins/*genetics/immunology/isolation & purification Molecular Sequence Dat; |
| Notes: | "MedlinePerin, M S Brose, N Jahn, R Sudhof, T C eng Comparative Study 1991/01/05 J Biol Chem. 1991 Jan 5; 266(1):623-9" |
