National Natural Science Foundation of China (NSFC)
31870724
中国
National Natural Science Foundation of China (NSFC)
82030108
中国
National Natural Science Foundation of China (NSFC)
31872796
中国
National Natural Science Foundation of China (NSFC)
81571127
中国
National Natural Science Foundation of China (NSFC)
31730006
中国
National Natural Science Foundation of China (NSFC)
31600606
中国
引用
ジャーナル: Cell Res / 年: 2022 タイトル: Structural basis of ALMT1-mediated aluminum resistance in Arabidopsis. 著者: Jiangqin Wang / Xiafei Yu / Zhong Jie Ding / Xiaokang Zhang / Yanping Luo / Ximing Xu / Yuan Xie / Xiaoxiao Li / Tian Yuan / Shao Jian Zheng / Wei Yang / Jiangtao Guo / 要旨: The plant aluminum (Al)-activated malate transporter ALMT1 mediates the efflux of malate to chelate the Al in acidic soils and underlies the plant Al resistance. Here we present cryo-electron ...The plant aluminum (Al)-activated malate transporter ALMT1 mediates the efflux of malate to chelate the Al in acidic soils and underlies the plant Al resistance. Here we present cryo-electron microscopy (cryo-EM) structures of Arabidopsis thaliana ALMT1 (AtALMT1) in the apo, malate-bound, and Al-bound states at neutral and/or acidic pH at up to 3.0 Å resolution. The AtALMT1 dimer assembles an anion channel and each subunit contains six transmembrane helices (TMs) and six cytosolic α-helices. Two pairs of Arg residues are located in the center of the channel pore and contribute to malate recognition. Al binds at the extracellular side of AtALMT1 and induces conformational changes of the TM1-2 loop and the TM5-6 loop, resulting in the opening of the extracellular gate. These structures, along with electrophysiological measurements, molecular dynamic simulations, and mutagenesis study in Arabidopsis, elucidate the structural basis for Al-activated malate transport by ALMT1.
凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277.15 K / 装置: FEI VITROBOT MARK IV
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
撮影
フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 平均露光時間: 8.0 sec. / 平均電子線量: 64.0 e/Å2
電子線
加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN
電子光学系
C2レンズ絞り径: 50.0 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm
実験機器
モデル: Titan Krios / 画像提供: FEI Company
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画像解析
粒子像選択
選択した数: 3586060 詳細: about 1,000 particles were manually picked from the micrographs for 2D classification initially. Class averages representing projections of AtALMT1 in different orientations were selected and ...詳細: about 1,000 particles were manually picked from the micrographs for 2D classification initially. Class averages representing projections of AtALMT1 in different orientations were selected and used as templates for automated particle picking
初期モデル
モデルのタイプ: OTHER 詳細: the ALMT1M60A/Al/pH5 model was built according to the reference ALMT1apo/pH5 model