介质载体15-kDa蛋白脂质与14-kDa多肽关联的证据。
Evidence for an association of the 15-kDa proteolipid of mediatophore with a 14-kDa polypeptide.
作者信息
Israël M, Morel N, Lesbats B
机构信息
Département de Neurochimie, CNRS, Gif-sur-Yvette, France.
出版信息
J Neurochem. 1991 Dec;57(6):2047-53. doi: 10.1111/j.1471-4159.1991.tb06420.x.
The present report shows that mediatophore, a nerve terminal membrane protein that translocates acetylcholine 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)dimethylammonio]-1-propanesulfonate 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, immunoprecipitation 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 synaptosomal membrane where mediatophore and its 15-kDa component are found.
本报告表明，介质载体是一种在钙作用下转运乙酰胆碱的神经终末膜蛋白，它与一种14 kDa的多肽形成复合物。该复合物是基于以下结果鉴定出来的。(a) 能免疫沉淀活性的多克隆抗介质载体抗血清沉淀出15 kDa和14 kDa的多肽。(b) 介质载体经高效液相色谱纯化后，两种抗原出现在同一峰中。(c) 用3-[(3-胆酰胺丙基)二甲基铵]-1-丙烷磺酸盐溶解突触前膜或纯化的介质载体后，用抗14 kDa抗原单克隆抗体制备的免疫亲和柱保留了14 kDa和15 kDa的多肽。同样，使用蛋白A包被的珠子进行免疫沉淀实验沉淀出一种免疫复合物，其中发现了两种抗原。(d) 14 kDa抗原可定位于突触体膜中，介质载体及其15 kDa成分也存在于该膜中。
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