纯化并重组的电鳐乙酰胆碱受体的膜片钳记录单通道电流。
Patch-recorded single-channel currents of the purified and reconstituted Torpedo acetylcholine receptor.
作者信息
Tank D W, Huganir R L, Greengard P, Webb W W
出版信息
Proc Natl Acad Sci U S A. 1983 Aug;80(16):5129-33. doi: 10.1073/pnas.80.16.5129.
Abstract
Small unilamellar vesicles containing purified and reconstituted nicotinic acetylcholine receptors from Torpedo electroplax have been fused by a simple freeze-thaw procedure to form large liposomes. Giga-seal patch-recording techniques were used to form isolated patches of liposome-membrane and to measure single-channel properties of the reconstituted receptor-ion channel complex. The observed properties are quantitatively similar to those reported for vertebrate muscle nicotinic acetylcholine receptor species recorded in situ. The results demonstrate that the pentameric complex consisting of the alpha 2 beta gamma delta subunits is fully functional. The methods used in these experiments should be useful in studying the effects of chemical alterations on the properties of acetylcholine receptor channels as well as other types of purified and reconstituted ion channels.
摘要
含有从电鳐电器官纯化并重组的烟碱型乙酰胆碱受体的小单层囊泡，通过简单的冻融程序融合形成大脂质体。采用千兆封接膜片钳记录技术形成脂质体膜的分离膜片，并测量重组受体 - 离子通道复合物的单通道特性。观察到的特性在数量上与原位记录的脊椎动物肌肉烟碱型乙酰胆碱受体种类所报道的特性相似。结果表明，由α2βγδ亚基组成的五聚体复合物具有完全功能。这些实验中使用的方法对于研究化学改变对乙酰胆碱受体通道以及其他类型纯化和重组离子通道特性的影响应该是有用的。
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本文引用的文献
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Acetylcholine receptor inactivation in Torpedo californica electroplax membrane vesicles. Detection of two processes in the millisecond and second time regions.加州电鳐电板膜囊泡中乙酰胆碱受体的失活。毫秒和秒时间区域内两个过程的检测。
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Mammalian muscle acetylcholine receptor: a supramolecular structure formed by four related proteins.哺乳动物肌肉乙酰胆碱受体：一种由四种相关蛋白质形成的超分子结构。
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The nicotinic cholinergic receptor: correlation of molecular structure with functional properties.烟碱型胆碱能受体：分子结构与功能特性的相关性
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Reconstitution of purified acetylcholine receptors with functional ion channels in planar lipid bilayers.在平面脂质双分子层中用功能性离子通道重组纯化的乙酰胆碱受体。
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Biophys J. 1983 Apr;42(1):1-10. doi: 10.1016/S0006-3495(83)84362-8.
10
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