Carboxy-terminal domain mediates assembly of the voltage-gated rat ether-a-go-go potassium channel
Carboxy-terminal domain mediates assembly of the voltage-gated rat ether-a-go-go potassium channel
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DOI：
10.1093/emboj/16.21.6337
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发表时间：
1997-11-03
期刊：
影响因子：
11.4
通讯作者：
Pongs, O
中科院分区：
文献类型：
--
作者：
Ludwig, J;Owen, D;Pongs, O
亚基与寡聚体的特异性组装是产生功能性钾通道的重要先决条件，我们通过两种互补测定研究了电压门控大鼠 ether-a-go-go (r-eag) 钾通道的组装，在蛋白质重叠结合实验中表明，靠近 r-eag 亚基羧基末端的 41 个氨基酸结构域对于 r-eag 亚基相互作用很重要，在体外表达系统中证明 r-eag 亚基缺乏该组装结构域不能形成功能性钾通道，此外，在共表达实验中，类似于10倍摩尔过量的r-eag羧基末端会抑制功能性r-eag通道的形成，当r-eag羧基末端组装结构域发生突变时，r-eag羧基末端对r-eag通道表达的显性负效应被消除。 r-eag 钾通道表达，与 Shaker 相关钾通道表达相反，Shaker 相关钾通道由氨基末端组装结构域引导。
The specific assembly of subunits to oligomers is an important prerequisite for producing functional potassium channels, We have studied the assembly of voltage-gated rat ether-a-go-go (r-eag) potassium channels with two complementary assays, In protein overlay binding experiments it was shown that a 41-amino-acid domain, close to the r-eag subunit carboxy-terminus, is important for r-eag subunit interaction, In an in vitro expression system it was demonstrated that r-eag subunits lacking this assembly domain cannot form functional potassium channels, Also, a similar to 10-fold molar excess of the r-eag carboxy-terminus inhibited in coexpression experiments the formation of functional r-eag channels, When the r-eag carboxy-terminal assembly domain had been mutated, the dominant-negative effect of the r-eag carboxy-terminus on r-eag channel expression was abolished, The results demonstrate that a carboxy-terminal assembly domain is essential for functional r-eag potassium channel expression, in contrast to the one of Shaker-related potassium channels, which is directed by an amino-terminal assembly domain.