Morales A, Aleu J, Ivorra I, Ferragut J A, Gonzalez-Ros J M, Miledi R
Department of Physiology, Universidad de Alicante, Spain.
Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8468-72. doi: 10.1073/pnas.92.18.8468.
Xenopus oocytes are a valuable aid for studying the molecular structure and function of ionic channels and neurotransmitter receptors. Their use has recently been extended by the demonstration that oocytes can incorporate foreign membranes carrying preassembled receptors and channels. Here we show that when reconstituted in an artificial lipid matrix and injected into Xenopus oocytes, purified nicotinic acetylcholine receptors are efficiently inserted into the plasma membrane, where they form "clusters" of receptors that retain their native properties. This constitutes an innovative approach that, besides allowing the analyses of membrane fusion processes, is also a powerful technique for studying the characteristics and regulation of many membrane proteins (with their native stoichiometry and configuration) upon reinsertion into the membrane of a very convenient host cell system.
非洲爪蟾卵母细胞对于研究离子通道和神经递质受体的分子结构与功能是一种很有价值的辅助工具。最近，卵母细胞能够整合携带预组装受体和通道的外源膜这一发现，拓展了其应用范围。在此我们表明，当在人工脂质基质中重构并注射到非洲爪蟾卵母细胞中时，纯化的烟碱型乙酰胆碱受体会有效地插入到质膜中，在那里它们形成保留其天然特性的受体“簇”。这构成了一种创新方法，除了能用于分析膜融合过程外，还是一种强大的技术，可用于研究许多膜蛋白（具有其天然化学计量和构型）重新插入到一个非常便利的宿主细胞系统的膜中后的特性和调控。
