Last T A, Gantzer M L, Tyler C D
J Biol Chem. 1983 Feb 25;258(4):2399-404.
An ion-gated channel was conferred on a planar lipid bilayer membrane upon incorporation of (Na+,K+)-ATPase. The channel exhibited two conductance states. The high conductance state was only observed when an ion gradient was present across the planar membrane. This state corresponded to an enzyme conformation which was ouabain and vanadate sensitive (i.e. conductance was inhibited by these compounds), while the low conductance state showed no sensitivity to either inhibitor. Single channel conductance behavior was observed when minimal amounts of enzyme were incorporated into the planar bilayer. The observed single channel conductance was 270 +/- 14 picosiemens. Similar transport behavior was observed for enzyme purified from ovine kidney using sodium dodecyl sulfate (anionic), eel electroplax using Lubrol-WX (nonionic), and kidney microsomes. In addition, the data strongly suggest that enzyme from the kidney microsomes was asymmetrically incorporated into the planar bilayer.
当将(钠,钾)-ATP酶整合到平面脂质双分子层膜上时,会赋予其一个离子门控通道。该通道呈现出两种电导状态。仅当平面膜两侧存在离子梯度时才会观察到高电导状态。这种状态对应于一种对哇巴因和钒酸盐敏感的酶构象(即电导被这些化合物抑制),而低电导状态对这两种抑制剂均不敏感。当将极少量的酶整合到平面双分子层中时,观察到了单通道电导行为。观察到的单通道电导为270±14皮西门子。使用十二烷基硫酸钠(阴离子型)从绵羊肾脏纯化的酶、使用Lubrol-WX(非离子型)从电鳗电板纯化的酶以及肾脏微粒体,都观察到了类似的转运行为。此外,数据强烈表明,来自肾脏微粒体的酶不对称地整合到了平面双分子层中。