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首页> 外文期刊>The Korean Journal of Physiology & Pharmacology >The Inhibition of TREK2 Channel by an Oxidizing Agent, 5,5'-dithio-bis (2-nitrobenzoic acid), via Interaction with the C-terminus Distal to the 353rd Amino Acid
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The Inhibition of TREK2 Channel by an Oxidizing Agent, 5,5'-dithio-bis (2-nitrobenzoic acid), via Interaction with the C-terminus Distal to the 353rd Amino Acid

机译：通过与C末端远端与353位氨基酸的相互作用，通过氧化剂5,5'-二硫代双（2-硝基苯甲酸）抑制TREK2通道

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摘要

TREK (TWIK-RElated K+ channels) and TRAAK (TWIK-Related Arachidonic acid Activated K+ channels) were expressed in COS-7 cells, and the channel activities were recorded from inside-out membrane patches using holding potential of -40 mV in symmetrical 150 mM K+ solution. Intracellular application of an oxidizing agent, 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB), markedly decreased the activity of the TREK2, and the activity was partially reversed by the reducing agent, dithiothreitol (DTT). In order to examine the possibility that the target sites for the oxidizing agents might be located in the C-terminus of TREK2, two chimeras were constructed: TREK2 (1-383)/TASK3C and TREK2 (1-353)/TASK3C. The channel activity in the TREK2 (1-383)/TASK3C chimera was still inhibited by DTNB, but not in the TREK2 (1-353)/TASK3C chimera. These results indicate that TREK2 is inhibited by oxidation, and that the target site for oxidation is located between the amino acid residues 353 and 383 in the C-terminus of the TREK2 protein.
机译：在COS-7细胞中表达TREK（TWIK相关的K + 通道）和TRAAK（TWIK相关的花生四烯酸激活的K + 通道），并记录其通道活性。在对称的150 mM K + 溶液中使用-40 mV的保持电位从内向外的膜片中分离。在细胞内使用氧化剂5,5'-二硫代双（2-硝基苯甲酸）（DTNB）会显着降低TREK2的活性，并且该活性会被还原剂二硫苏糖醇（DTT）部分逆转。为了检查氧化剂的目标位点可能位于TREK2的C端的可能性，构建了两种嵌合体：TREK2（1-383）/ TASK3C和TREK2（1-353）/ TASK3C。 DTNB仍然抑制TREK2（1-383）/ TASK3C嵌合体中的通道活性，但不抑制TREK2（1-353）/ TASK3C嵌合体中的通道活性。这些结果表明TREK2被氧化抑制，并且氧化的靶位点位于TREK2蛋白的C末端的氨基酸残基353和383之间。

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