Molecular mechanism underlying regulation of Arabidopsis CLCa transporter by nucleotides and phospholipids.
Molecular mechanism underlying regulation of Arabidopsis CLCa transporter by nucleotides and phospholipids.
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DOI：
10.1038/s41467-023-40624-z
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发表时间：
2023-08-12
影响因子：
16.6
通讯作者：
Zhang, Peng
中科院分区：
文献类型：
--
作者：
Yang, Zhao;Zhang, Xue;Ye, Shiwei;Zheng, Jingtao;Huang, Xiaowei;Yu, Fang;Chen, Zhenguo;Cai, Shiqing;Zhang, Peng
氯离子通道（Chloride channels，CLC）是一种跨膜转运阴离子的通道，可调节细胞内离子平衡和细胞器的酸化，分为阴离子通道和阴离子/质子反向转运体。拟南芥CLCa（AtCLCa）转运蛋白定位于液泡膜，从细胞质中输入NO3−和较小程度的Cl−。AtCLCa和许多其他CLC的活性受核苷酸和磷脂的调节，但其分子机制尚不清楚。在这里，我们确定的cryo-EM结构AtCLCa绑定NO3−和Cl−，分别。这两种结构都以ATP和PI（4，5）P2结合构象捕获。结构和电生理学分析揭示了先前未鉴定的N-末端β-发夹，其通过ATP结合稳定以阻断阴离子转运途径，从而抑制AtCLCa活性。而AMP由于缺乏β-发夹稳定所需的β/γ-磷酸而失去抑制能力。这很好地解释了AtCLCa如何感知ATP/AMP状态来调节生理氮碳平衡。我们的数据进一步表明PI（4，5）P2或PI（3，5）P2结合到AtCLCa二聚体界面并占据质子出口途径，这可能有助于理解磷脂对AtCLCa的抑制，以促进保卫细胞液泡酸化和气孔关闭。总之，我们的工作揭示了在特定的生理条件下，核苷酸和磷脂对AtCLCa的调控机制，为进一步研究CLCs提供了新的思路。CLC转运蛋白受核苷酸和磷脂调节。在这里，拟南芥CLCa与ATP和PIP 2复合的冷冻电镜结构和电生理分析表明了核苷酸和磷脂的潜在调控机制。
Chloride channels (CLCs) transport anion across membrane to regulate ion homeostasis and acidification of intracellular organelles, and are divided into anion channels and anion/proton antiporters. Arabidopsis thaliana CLCa (AtCLCa) transporter localizes to the tonoplast which imports NO3− and to a less extent Cl− from cytoplasm. The activity of AtCLCa and many other CLCs is regulated by nucleotides and phospholipids, however, the molecular mechanism remains unclear. Here we determine the cryo-EM structures of AtCLCa bound with NO3− and Cl−, respectively. Both structures are captured in ATP and PI(4,5)P2 bound conformation. Structural and electrophysiological analyses reveal a previously unidentified N-terminal β-hairpin that is stabilized by ATP binding to block the anion transport pathway, thereby inhibiting the AtCLCa activity. While AMP loses the inhibition capacity due to lack of the β/γ- phosphates required for β-hairpin stabilization. This well explains how AtCLCa senses the ATP/AMP status to regulate the physiological nitrogen-carbon balance. Our data further show that PI(4,5)P2 or PI(3,5)P2 binds to the AtCLCa dimer interface and occupies the proton-exit pathway, which may help to understand the inhibition of AtCLCa by phospholipids to facilitate guard cell vacuole acidification and stomatal closure. In a word, our work suggests the regulatory mechanism of AtCLCa by nucleotides and phospholipids under certain physiological scenarios and provides new insights for future study of CLCs. CLC transporters are regulated by nucleotides and phospholipids. Here cryo-EM structure of Arabidopsis CLCa in complex with ATP and PIP2 and electrophysiological analysis suggests the underlying regulatory mechanisms of both nucleotides and phospholipids.
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影响因子：
3.8
作者：
Accardi, Alessio;Walden, Michael;Nguitragool, Wang;Jayaram, Hariharan;Williams, Carole;Miller, Christopher
通讯作者：
Miller, Christopher
影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
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：
64.8
作者：
Graves, Austin R.;Curran, Patricia K.;Mindell, Joseph A.
通讯作者：
Mindell, Joseph A.
影响因子：
56.9
作者：
Dutzler, R;Campbell, EB;MacKinnon, R
通讯作者：
MacKinnon, R