Kim M, Baro D J, Lanning C C, Doshi M, Farnham J, Moskowitz H S, Peck J H, Olivera B M, Harris-Warrick R M
Section of Neurobiology and Behavior, Cornell University, Ithaca, New York 14853, USA.
J Neurosci. 1997 Nov 1;17(21):8213-24. doi: 10.1523/JNEUROSCI.17-21-08213.1997.
We have cloned cDNAs for the shaker potassium channel gene from the spiny lobster Panulirus interruptus. As previously found in Drosophila, there is alternative splicing at the 5' and 3' ends of the coding region. However, in Panulirus shaker, alternative splicing also occurs within the pore-forming region of the protein. Three different splice variants were found within the P region, two of which bestow unique electrophysiological characteristics to channel function. Pore I and pore II variants differ in voltage dependence for activation, kinetics of inactivation, current rectification, and drug resistance. The pore 0 variant lacks a P region exon and does not produce a functional channel. This is the first example of alternative splicing within the pore-forming region of a voltage-dependent ion channel. We used a recently identified potassium channel blocker, kappa-conotoxin PVIIA, to study the physiological role of the two pore forms. The toxin selectively blocked one pore form, whereas the other form, heteromers between the two pore forms, and Panulirus shal were not blocked. When it was tested in the Panulirus stomatogastric ganglion, the toxin produced no effects on transient K+ currents or synaptic transmission between neurons.
我们已经克隆了来自多棘龙虾(Panulirus interruptus)的震颤钾通道基因的cDNA。正如之前在果蝇中发现的那样,在编码区的5'端和3'端存在可变剪接。然而,在多棘龙虾的震颤通道中,可变剪接也发生在蛋白质的孔形成区域内。在P区域内发现了三种不同的剪接变体,其中两种赋予通道功能独特的电生理特性。孔I和孔II变体在激活的电压依赖性、失活动力学、电流整流和耐药性方面存在差异。孔0变体缺少一个P区域外显子,不产生功能性通道。这是电压依赖性离子通道孔形成区域内可变剪接的第一个例子。我们使用最近鉴定出的一种钾通道阻滞剂——κ-芋螺毒素PVIIA,来研究这两种孔形式的生理作用。该毒素选择性地阻断一种孔形式,而另一种孔形式、两种孔形式之间的异源二聚体以及多棘龙虾的shal通道均未被阻断。当在多棘龙虾的口胃神经节中进行测试时,该毒素对瞬态K+电流或神经元之间的突触传递没有影响。