Variomics Screen Identifies the Re-entrant Loop of the Calcium-activated Chloride Channel ANO1 That Facilitates Channel Activation
Variomics Screen Identifies the Re-entrant Loop of the Calcium-activated Chloride Channel ANO1 That Facilitates Channel Activation
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DOI：
10.1074/jbc.m114.618140
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发表时间：
2015-01-09
影响因子：
4.8
通讯作者：
Gaither, L. Alex
中科院分区：
文献类型：
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作者：
Bill, Anke;Popa, M. Oana;Gaither, L. Alex
钙激活的氯离子通道ANO 1调节多种生理过程。然而，对通道门控和ANO 1活性调节的机制知之甚少。使用高通量，随机诱变为基础的变异组学筛选，我们产生和功能特征类似于6000 ANO 1突变体，并确定了新的突变，影响通道活性，细胞内运输，或本地化的ANO 1。突变如S741 T增加了ANO 1钙敏感性，并使ANO 1钙门控电压无关，证明了折返环在耦合ANO 1的钙和电压敏感性中的关键作用，因此在调节ANO 1激活中。我们的数据提出了第一个公正的和全面的研究的结构-功能关系的ANO 1。新的ANO 1突变体具有多种功能特征，为研究ANO 1在生物系统中的功能提供了新的工具，为更好地了解ANO 1的功能及其在健康和疾病中的作用铺平了道路。
The calcium-activated chloride channel ANO1 regulates multiple physiological processes. However, little is known about the mechanism of channel gating and regulation of ANO1 activity. Using a high-throughput, random mutagenesis-based variomics screen, we generated and functionally characterized similar to 6000 ANO1 mutants and identified novel mutations that affected channel activity, intracellular trafficking, or localization of ANO1. Mutations such as S741T increased ANO1 calcium sensitivity and rendered ANO1 calcium gating voltage-independent, demonstrating a critical role of the re-entrant loop in coupling calcium and voltage sensitivity of ANO1 and hence in regulating ANO1 activation. Our data present the first unbiased and comprehensive study of the structure-function relationship of ANO1. The novel ANO1 mutants reported have diverse functional characteristics, providing new tools to study ANO1 function in biological systems, paving the path for a better understanding of the function of ANO1 and its role in health and diseases.