Grissmer S, Lewis R S, Cahalan M D
Department of Physiology and Biophysics, University of California, Irvine 92717.
J Gen Physiol. 1992 Jan;99(1):63-84. doi: 10.1085/jgp.99.1.63.
Using the patch-clamp technique, we have identified two types of Ca(2+)-activated K+ (K(Ca)) channels in the human leukemic T cell line. Jurkat. Substances that elevate the intracellular Ca2+ concentration ([Ca2+]i), such as ionomycin or the mitogenic lectin phytohemagglutinin (PHA), as well as whole-cell dialysis with pipette solutions containing elevated [Ca2+]i, activate a voltage-independent K+ conductance. Unlike the voltage-gated (type n) K+ channels in these cells, the majority of K(Ca) channels are insensitive to block by charybdotoxin (CTX) or 4-aminopyridine (4-AP), but are highly sensitive to block by apamin (Kd less than 1 nM). Channel activity is strongly dependent on [Ca2+]i, suggesting that multiple Ca2+ binding sites may be involved in channel opening. The Ca2+ concentration at which half of the channels are activated is 400 nM. These channels show little voltage dependence over a potential range of -100 to 0 mV and have a unitary conductance of 4-7 pS in symmetrical 170 mM K+. In the presence of 10 nM apamin, a less prevalent type of K(Ca) channel with a unitary conductance of 40-60 pS can be observed. These larger-conductance channels are sensitive to block by CTX. Pharmacological blockade of K(Ca) channels and voltage-gated type n channels inhibits oscillatory Ca2+ signaling triggered by PHA. These results suggest that K(Ca) channels play a supporting role during T cell activation by sustaining dynamic patterns of Ca2+ signaling.
运用膜片钳技术,我们在人白血病T细胞系Jurkat中鉴定出了两种类型的钙激活钾(K(Ca))通道。能提高细胞内钙离子浓度([Ca2+]i)的物质,如离子霉素或促有丝分裂凝集素植物血凝素(PHA),以及用含有升高的[Ca2+]i的移液管溶液进行全细胞透析,均可激活一种电压非依赖性钾电导。与这些细胞中的电压门控(n型)钾通道不同,大多数K(Ca)通道对蝎毒素(CTX)或4-氨基吡啶(4-AP)的阻断不敏感,但对蜂毒明肽的阻断高度敏感(解离常数小于1 nM)。通道活性强烈依赖于[Ca2+]i,这表明多个钙离子结合位点可能参与通道开放。半数通道被激活时的钙离子浓度为400 nM。这些通道在-100至0 mV的电位范围内几乎没有电压依赖性,在对称的170 mM K+中单位电导为4 - 7 pS。在存在10 nM蜂毒明肽的情况下,可以观察到一种不太常见的K(Ca)通道,其单位电导为40 - 60 pS。这些较大电导的通道对CTX的阻断敏感。K(Ca)通道和电压门控n型通道的药理学阻断可抑制由PHA触发的振荡性钙离子信号传导。这些结果表明,K(Ca)通道在T细胞激活过程中通过维持钙离子信号传导的动态模式发挥辅助作用。