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培养的大鼠肾上腺嗜铬细胞中小电导钙激活钾通道的离子选择性和门控特性

Ion selectivity and gating of small conductance Ca(2+)-activated K+ channels in cultured rat adrenal chromaffin cells.

作者信息

Park Y B

机构信息

Department of Physiology and Biophysics, University of Washington, School of Medicine, Seattle 98195.

出版信息

J Physiol. 1994 Dec 15;481 ( Pt 3)(Pt 3):555-70. doi: 10.1113/jphysiol.1994.sp020463.

Abstract
  1. The ion selectivity and gating of apamin-sensitive, small conductance Ca(2+)-activated K+ (SK) channels were studied in cultured rat adrenal chromaffin cells using patch clamp techniques. 2. The amplitude of slow tail currents showed a bell-shaped dependence on depolarization potentials. Slow tail currents were abolished in a Ca(2+)-free external solution or by adding 100 microM Cd2+ to the external solution. Reversal potentials followed the predictions of the Nernst equation for a K+ electrode. 3. Slow tail currents were largely blocked by external application of apamin (dissociation constant, Kd, 4.4 nM), (+)-tubocurarine (Kd, 20 microM), and tetraethylammonium (Kd, 5.4 mM). 4. The relative permeability (PX/PK, where X may be any one of the ions listed) of SK channels was: Tl+ (1.87) > K+ (1.0) > Rb+ (0.81) > Cs+ (0.16) > NH4+ (0.11). Na+, Li+ and methylamine were not measurably permeant (PX/PK < 0.005). Open SK channels seem to have an effective pore diameter of 0.34-0.38 nm. The relative conductance (gX/gK) was: Tl+ (1.29) > K+ (1.0) > Rb+ (0.85) > Cs+ (0.45) approximately NH4+ (0.44). 5. With mixtures of Tl+ and K+, SK channels showed anomalous mole-fraction behaviour. 6. Ca2+ dependence of SK channel gating was studied using inside-out macropatches. The [Ca2+] required for half-maximal activation and the Hill coefficient were 0.69 microM and 1.7, respectively, and independent of membrane potentials. 7. Single-channel conductance was 13-14 pS (160 mM K+).
摘要
  1. 运用膜片钳技术,在培养的大鼠肾上腺嗜铬细胞中研究了蜂毒明肽敏感的小电导钙激活钾(SK)通道的离子选择性和门控特性。2. 慢尾电流的幅度对去极化电位呈钟形依赖性。在无钙的外部溶液中或向外部溶液中添加100μM Cd2+时,慢尾电流消失。反转电位符合钾电极的能斯特方程预测。3. 外部施加蜂毒明肽(解离常数,Kd,4.4 nM)、(+) - 筒箭毒碱(Kd,20μM)和四乙铵(Kd,5.4 mM)可很大程度上阻断慢尾电流。4. SK通道的相对通透性(PX/PK,其中X可以是所列的任何一种离子)为:Tl+(1.87)> K+(1.0)> Rb+(0.81)> Cs+(0.16)> NH4+(0.11)。Na+、Li+和甲胺的通透性不可测量(PX/PK < 0.005)。开放的SK通道似乎具有0.34 - 0.38 nm的有效孔径。相对电导率(gX/gK)为:Tl+(1.29)> K+(1.0)> Rb+(0.85)> Cs+(0.45)≈NH4+(0.44)。5. 对于Tl+和K+的混合物,SK通道表现出异常的摩尔分数行为。6. 使用内面向外的大膜片研究了SK通道门控的钙依赖性。半最大激活所需的[Ca2+]和希尔系数分别为0.69μM和1.7,且与膜电位无关。7. 单通道电导为13 - 14 pS(160 mM K+)。

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