从电鳐电器官大规模纯化突触前质膜。
Large-scale purification of presynaptic plasma membranes from Torpedo marmorata electric organ.
作者信息
Morel N, Marsal J, Manaranche R, Lazereg S, Mazie J C, Israel M
出版信息
J Cell Biol. 1985 Nov;101(5 Pt 1):1757-62. doi: 10.1083/jcb.101.5.1757.
Abstract
The presynaptic plasma membrane (PSPM) of cholinergic nerve terminals was purified from Torpedo electric organ using a large-scale procedure. Up to 500 g of frozen electric organ were fractioned in a single run, leading to the isolation of greater than 100 mg of PSPM proteins. The purity of the fraction is similar to that of the synaptosomal plasma membrane obtained after subfractionation of Torpedo synaptosomes as judged by its membrane-bound acetylcholinesterase activity, the number of Glycera convoluta neurotoxin binding sites, and the binding of two monoclonal antibodies directed against PSPM. The specificity of these antibodies for the PSPM is demonstrated by immunofluorescence microscopy.
摘要
利用大规模方法从电鳐电器官中纯化出胆碱能神经末梢的突触前质膜（PSPM）。单次操作可对多达500克冷冻电器官进行分级分离，从而分离出超过100毫克的PSPM蛋白。通过其膜结合乙酰胆碱酯酶活性、甘油涡虫神经毒素结合位点数量以及两种针对PSPM的单克隆抗体的结合情况判断，该分级分离物的纯度与电鳐突触体亚分级分离后获得的突触体质膜的纯度相似。免疫荧光显微镜证实了这些抗体对PSPM的特异性。
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