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首页> 外文期刊>The European Journal of Neuroscience >Inhibition of BK channel activity by association with calcineurin in rat brain.
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Inhibition of BK channel activity by association with calcineurin in rat brain.

机译:与钙调神经磷酸酶联结抑制BK通道活性在大鼠脑中。

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Large conductance calcium-activated potassium (BK(Ca)) channels are regulated by a number of different protein kinases and phosphatases. The close association of enzymes and channel have been shown to underlie many examples of modulation. However, only the association of protein kinase A with the BK(Ca) channel has been detailed [Tian et al. (2003)J. Biol. Chem., 278, 8669-8677]. We have found using reciprocal immunoprecipitations that the BK(Ca) channel associates with the calcium/calmodulin-dependent phosphatase calcineurin, in Wistar rat brain. A HA-tagged construct of the carboxyl terminus of rSlo(27), a variant of the BK(Ca) channel that is abundant in the hippocampus [Ha et al. (2000)Eur. J. Biochem., 267, 910-9218], was found to associate only with the B subunit of calcineurin. This data suggests that the majority of the interaction of the BK(Ca) channel with calcineurin is mediated by the B subunit of the phosphatase. This was confirmed by using glutathione-S-transferase (GST) fusion proteins of the linker regions between the S7-S10 hydrophobic domains in the carboxyl terminus of rSlo(27), where only the B subunit of calcineurin interacted with regions between S7 and S9 of the channel. Addition of a constitutively active calcineurin (CaN(420)) to inside-out membrane patches excised from cultured hippocampal neurons resulted in a dramatic reduction in BK(Ca) channel open probability, with only very short-duration events being apparent. These data suggest that BK(Ca) channel activity is inhibited by calcineurin, an effect mediated by the association of the calcineurin B subunit with the carboxyl terminus of the channel.
机译:大电导钙激活钾(BK(Ca))通道受许多不同的蛋白激酶和磷酸酶调节。酶和通道的紧密结合已被证明是许多调节的基础。但是,只有蛋白激酶A与BK(Ca)通道的关联才被详细描述[Tian等。 (2003)J.生物学278. 8669-8677]。我们已经发现在Wistar大鼠脑中使用相互免疫沉淀BK(Ca)通道与钙/钙调蛋白依赖性磷酸酶钙调神经磷酸酶相关。 HA标记的rSlo(27)羧基末端的构建体,rSlo(27)是海马中丰富的BK(Ca)通道的变异体[Ha等。 (2000)欧元。 [J. Biochem。,267,910-9218]被发现仅与钙调神经磷酸酶的B亚基缔合。该数据表明,BK(Ca)通道与钙调神经磷酸酶的大部分相互作用是由磷酸酶的B亚基介导的。这是通过使用rSlo(27)羧基末端S7-S10疏水域之间的接头区域的谷胱甘肽S-转移酶(GST)融合蛋白来证实的,其中仅钙调神经磷酸酶的B亚基与S7和S9之间的区域相互作用频道的向从培养的海马神经元切出的内而外膜片中添加组成型活性钙调神经磷酸酶(CaN(420))会导致BK(Ca)通道开放可能性大大降低,只有很短的持续时间才明显。这些数据表明钙调神经磷酸酶抑制了BK(Ca)通道的活性,钙调神经磷酸酶B亚基与通道的羧基末端之间的缔合介导了这种作用。

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