Labarca P P, Miller C
J Membr Biol. 1981;61(1):31-8. doi: 10.1007/BF01870750.
Sarcoplasmic reticulum (SR) vesicles from frog leg muscle were fused with a planar phospholipid bilayer by a method described previously for rabbit SR. As a result of the fusion, K+-selective conduction channels are inserted into the bilayer. Unlike the two-state rabbit channel, the frog channel displays three states: a nonconducting ("closed") state and two conducting states "alpha" and "beta". In 0.1 M K+ the single-channel conductances are 50 and 150 pS for alpha and beta, respectively. The probabilities of appearance of the three states are voltage-dependent, and transitions between the closed and beta states proceed through the alpha state. Both open states follow a quantitatively identical selectivity sequence in channel conductance: K+ greater than NH4+ greater than Rb+ greater than Na+ greater than Li+ greater than Cs+. Both open states are blocked by Cs+ asymmetrically in a voltage-dependent manner. The zero-voltage dissociation constant for blocking is the same for both open states, but the voltage-dependences of the Cs+ block for the two states differ in a way suggesting that the Cs+ blocking site is located more deeply inside the membrane in the beta than in the alpha state.
通过先前描述的用于兔肌浆网(SR)的方法,将来自蛙腿肌肉的肌浆网(SR)囊泡与平面磷脂双层融合。融合的结果是,钾离子选择性传导通道插入到双层中。与双态的兔通道不同,蛙通道呈现三种状态:一种非传导性(“关闭”)状态和两种传导状态“α”和“β”。在0.1M钾离子溶液中,α态和β态的单通道电导分别为50和150皮西门子。这三种状态出现的概率取决于电压,并且在关闭态和β态之间的转变通过α态进行。两种开放状态在通道电导方面遵循定量相同的选择性序列:钾离子>铵根离子>铷离子>钠离子>锂离子>铯离子。两种开放状态都被铯离子以电压依赖性方式不对称地阻断。两种开放状态的零电压阻断解离常数相同,但两种状态下铯离子阻断的电压依赖性不同,这表明β态下铯离子阻断位点比α态更深入膜内。