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Evidence for an Association of the 15‐kDa Proteolipid of Mediatophore with a 14‐kDa Polypeptide
Author(s) -
Israël M.,
Morel N.,
Lesbats B.
Publication year - 1991
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1991.tb06420.x
Subject(s) - polyclonal antibodies , antiserum , antigen , monoclonal antibody , biochemistry , molecular mass , immunoprecipitation , membrane , microbiology and biotechnology , biology , membrane protein , affinity chromatography , chemistry , antibody , enzyme , gene , genetics , immunology
The present report shows that mediatophore, a nerve terminal membrane protein that translocates acetyl‐choline on calcium action, forms a complex with a 14‐kDa polypeptide. The complex was identified based on the following results, (a) A polyclonal antimediatophore antiserum that immunoprecipitates activity precipitates both the 15‐and 14‐kDa polypeptides. (b) After HPLC purification of mediatophore, both antigens were found in the same peak, (c) After 3‐[(3‐cholamidopropyl)dimethylarnmonio]‐1‐pro‐panesulfonate solubilization of presynaptic membranes or of the purified mediatophore, an immunoaffinity column made with the anti‐14‐kDa antigen monoclonal antibody retained both the 14‐kDa and the 15‐kDa polypeptide. Similarly, im‐munoprecipitation experiments using protein A‐coated beads sedimented an immunocomplex in which both antigens were found, (d) The 14‐kDa antigen could be localized in the syn‐aptosomal membrane where mediatophore and its 15‐kDa component are found.