Shenkel S, Cooper E C, James W, Agnew W S, Sigworth F J
Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06510.
Proc Natl Acad Sci U S A. 1989 Dec;86(23):9592-6. doi: 10.1073/pnas.86.23.9592.
A recent study showed that limited trypsin treatment of liposomes containing purified Electrophorus electricus sodium channels activates a sodium radiotracer flux. We now report that similarly treated sodium channels show voltage-gated, tetrodotoxin-sensitive and highly sodium-selective single-channel currents when incorporated into planar lipid membranes. The trypsinized channels opened repeatedly in bursts of several seconds duration, as would be expected for channels whose fast inactivation process had been removed. Furthermore, they have a higher conductance, different voltage-dependence of gating, and a remarkably higher selectivity (PNa/PK = 41) than sodium channels bound by batrachotoxin or other activating neurotoxins; these properties of the trypsinized channels are probably closer to those of channels in intact electrocytes.
最近的一项研究表明,用有限量的胰蛋白酶处理含有纯化的电鳗钠通道的脂质体,可激活钠放射性示踪剂通量。我们现在报告,当整合到平面脂质膜中时,经过类似处理的钠通道表现出电压门控、河豚毒素敏感且高度钠选择性的单通道电流。经胰蛋白酶处理的通道会以持续数秒的爆发形式反复打开,这与快速失活过程已被去除的通道预期情况相符。此外,与被蟾毒素或其他激活神经毒素结合的钠通道相比,它们具有更高的电导、不同的门控电压依赖性以及显著更高的选择性(PNa/PK = 41);经胰蛋白酶处理的通道的这些特性可能更接近完整电细胞中通道的特性。