Zhang X, Ma J, Berkowitz G A
Department of Plant Science, University of Connecticut, Storrs, Connecticut 06269-4067, USA.
Plant Physiol. 1999 Nov;121(3):995-1002. doi: 10.1104/pp.121.3.995.
Animal K(+) channel alpha- (pore-forming) subunits form native proteins by association with beta-subunits, which are thought to affect channel function by modifying electrophysiological parameters of currents (often by inducing fast inactivation) or by stabilizing the protein complex. We evaluated the functional association of KAT1, a plant K(+) channel alpha-subunit, and KAB1 (a putative homolog of animal K(+) channel beta-subunits) by co-expression in Xenopus laevis oocytes. Oocytes expressing KAT1 displayed inward-rectifying, non-inactivating K(+) currents that were similar in magnitude to those reported in prior studies. K(+) currents recorded from oocytes expressing both KAT1 and KAB1 had similar gating kinetics. However, co-expression resulted in greater total current, consistent with the possibility that KAB1 is a beta-subunit that stabilizes and therefore enhances surface expression of K(+) channel protein complexes formed by alpha-subunits such as KAT1. K(+) channel protein complexes formed by alpha-subunits such as KAT1 that undergo (voltage-dependent) inactivation do so by means of a "ball and chain" mechanism; the ball portion of the protein complex (which can be formed by the N terminus of either an alpha- or beta-subunit) occludes the channel pore. KAT1 was co-expressed in oocytes with an animal K(+) channel alpha-subunit (hKv1.4) known to contain the N-terminal ball and chain. Inward currents through heteromeric hKv1. 4:KAT1 channels did undergo typical voltage-dependent inactivation. These results suggest that inward currents through K(+) channel proteins formed at least in part by KAT1 polypeptides are capable of inactivation, but the structural component facilitating inactivation is not present when channel complexes are formed by either KAT1 or KAB1 in the absence of additional subunits.
动物钾离子通道α(形成孔道的)亚基通过与β亚基结合形成天然蛋白质,β亚基被认为通过改变电流的电生理参数(通常是诱导快速失活)或稳定蛋白质复合物来影响通道功能。我们通过在非洲爪蟾卵母细胞中共表达,评估了植物钾离子通道α亚基KAT1与KAB1(动物钾离子通道β亚基的推定同源物)的功能关联。表达KAT1的卵母细胞表现出内向整流、非失活的钾离子电流,其大小与先前研究报道的相似。从同时表达KAT1和KAB1的卵母细胞记录到的钾离子电流具有相似的门控动力学。然而,共表达导致总电流更大,这与KAB1是一种β亚基的可能性一致,该β亚基可稳定并因此增强由α亚基(如KAT1)形成的钾离子通道蛋白复合物的表面表达。由α亚基(如KAT1)形成的钾离子通道蛋白复合物通过“球与链”机制发生(电压依赖性)失活;蛋白质复合物的球部分(可由α或β亚基的N端形成)堵塞通道孔。KAT1与已知含有N端球与链的动物钾离子通道α亚基(hKv1.4)在卵母细胞中共表达。通过异源hKv1.4:KAT1通道的内向电流确实经历了典型的电压依赖性失活。这些结果表明,至少部分由KAT1多肽形成的钾离子通道蛋白的内向电流能够失活,但在没有其他亚基的情况下,当通道复合物由KAT1或KAB1形成时,促进失活的结构成分不存在。