Delcour A H, Martinac B, Adler J, Kung C
Department of Biochemistry, University of Wisconsin, Madison 53706.
Biophys J. 1989 Sep;56(3):631-6. doi: 10.1016/S0006-3495(89)82710-9.
We have modified the procedure of Criado and Keller (1987) to study ion channels of Escherichia coli reconstituted in liposomes. The modifications include (a) excluding the use of any detergent and (b) inducing blisters from liposomes with Mg2+. These blisters, which appear to be unilamellar, are stable for hours. They could be repeatedly sampled with different patch-clamp pipettes each achieving seal resistance greater than 10 GOhms. Activities of three types of ion channels are often observed by use of this method, including two voltage-sensitive cation channels of different conductances. Even the mechanosensitive channel, previously recorded from live E. coli cells (Martinac et al., 1987), was also detected in these blisters. Apparently the channel protein and any accessory structures, postulated to be needed for mechanotransduction, can be reconstituted together by this method.
我们修改了Criado和Keller（1987年）的实验步骤，以研究脂质体中重组的大肠杆菌离子通道。修改内容包括：（a）不使用任何去污剂；（b）用Mg2+诱导脂质体形成水泡。这些水泡似乎是单层的，能稳定存在数小时。可以用不同的膜片钳吸管反复取样，每个吸管的封接电阻都大于10 GΩ。使用这种方法经常能观察到三种类型离子通道的活性，包括两种不同电导的电压敏感阳离子通道。甚至之前在活的大肠杆菌细胞中记录到的机械敏感通道（Martinac等人，1987年），在这些水泡中也被检测到。显然，这种方法可以将推测为机械转导所需的通道蛋白和任何辅助结构一起重组。
