Hazelbauer G L, Changeux J P
Proc Natl Acad Sci U S A. 1974 Apr;71(4):1479-83. doi: 10.1073/pnas.71.4.1479.
Membrane fragments rich in cholinergic (nicotinic) receptor sites, purified from homogenates of Torpedo marmorata electric organ, are dissolved in high concentrations of Na cholate and Tris buffer in the presence of Na ethylenediaminetetraacetate, without loss of the ability to bind Naja nigricollis [(3)H]alpha-toxin. After extensive dialysis to remove cholate, the solution is supplemented with crude lipids extracted from native membrane fragments and MgCl(2)-CaCl(2) added dropwise to the mixture as a concentrated solution. The reconstituted membranes obtained retain (22)Na(+). Release of Na from these microsacs increases in the presence of carbamylcholine and the alpha-toxin from N. nigricollis blocks this effect.
从电鳐电器官匀浆中纯化得到的富含胆碱能(烟碱型)受体位点的膜片段,在乙二胺四乙酸钠存在的情况下,溶解于高浓度的胆酸钠和 Tris 缓冲液中,且不丧失结合眼镜蛇毒[(3)H]α-毒素的能力。经过广泛透析以去除胆酸盐后,向溶液中补充从天然膜片段中提取的粗脂质,并将氯化镁-氯化钙作为浓缩溶液逐滴加入混合物中。所获得的重构膜保留了(22)Na(+)。在氨甲酰胆碱存在的情况下,这些微囊中的钠释放增加,而来自黑颈眼镜蛇的α-毒素可阻断这种效应。