Cryo-EM structures of ClC-2 chloride channel reveal the blocking mechanism of its specific inhibitor AK-42.
Cryo-EM structures of ClC-2 chloride channel reveal the blocking mechanism of its specific inhibitor AK-42.
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DOI：
10.1038/s41467-023-39218-6
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发表时间：
2023-06-09
影响因子：
16.6
通讯作者：
Zhang, Huawei
中科院分区：
文献类型：
--
作者：
Ma, Tao;Wang, Lei;Chai, Anping;Liu, Chao;Cui, Wenqiang;Yuan, Shuguang;Au, Shannon Wing Ngor;Sun, Liang;Zhang, Xiaokang;Zhang, Zhenzhen;Lu, Jianping;Gao, Yuanzhu;Wang, Peiyi;Li, Zhifang;Liang, Yujie;Vogel, Horst;Wang, Yu Tian;Wang, Daping;Yan, Kaige;Zhang, Huawei
ClC-2转运氯离子穿过质膜，在细胞内稳态中起关键作用。它的功能障碍与包括脑白质营养不良和原发性醛固酮增多症在内的疾病有关。AK-42最近被报道为ClC-2的特异性抑制剂。然而，实验结构仍然缺乏破译其抑制机制。在这里，我们提出了cryo-EM结构的载脂蛋白ClC-2和它的复合物与AK-42，都在3.5毫米的分辨率。残基S162、E205和Y553参与氯离子结合并有助于离子选择性。门控谷氨酸E205的侧链占据了假定的中央氯离子结合位点，表明我们的结构代表了封闭状态。结构分析、分子动力学和电生理记录确定了与AK-42相互作用的关键残基。几个AK-42相互作用的残基存在于ClC-2中，但不存在于其他ClC中，为AK-42特异性提供了可能的解释。综上所述，我们的实验结果揭示了ClC-2抑制剂AK-42的潜在抑制机制。AK-42被报道为ClC-2的特异性抑制剂，但其作用机制尚不清楚。在这里，作者报告了载脂蛋白ClC-2和AK-42复合物的冷冻电镜结构，揭示了它如何特异性抑制ClC-2。
ClC-2 transports chloride ions across plasma membranes and plays critical roles in cellular homeostasis. Its dysfunction is involved in diseases including leukodystrophy and primary aldosteronism. AK-42 was recently reported as a specific inhibitor of ClC-2. However, experimental structures are still missing to decipher its inhibition mechanism. Here, we present cryo-EM structures of apo ClC-2 and its complex with AK-42, both at 3.5 Å resolution. Residues S162, E205 and Y553 are involved in chloride binding and contribute to the ion selectivity. The side-chain of the gating glutamate E205 occupies the putative central chloride-binding site, indicating that our structure represents a closed state. Structural analysis, molecular dynamics and electrophysiological recordings identify key residues to interact with AK-42. Several AK-42 interacting residues are present in ClC-2 but not in other ClCs, providing a possible explanation for AK-42 specificity. Taken together, our results experimentally reveal the potential inhibition mechanism of ClC-2 inhibitor AK-42. AK-42 was reported as a specific inhibitor of ClC-2 but its working mechanism is not clear. Here authors report cryo-EM structures of apo ClC-2 and in complex with AK-42, revealing how it inhibits ClC-2 specifically.
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影响因子：
5.8
作者：
Gouet, P;Courcelle, E;Métoz, F
通讯作者：
Métoz, F
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
3.8
作者：
De Jesus-Perez, Jose J.;Castro-Chong, Alejandra;Arreola, Jorge
通讯作者：
Arreola, Jorge
影响因子：
56.9
作者：
Dutzler, R;Campbell, EB;MacKinnon, R
通讯作者：
MacKinnon, R
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH