Netherlands Organisation for Scientific Research (NWO)
722.017.001
オランダ
Norwegian Research Council
234817
ノルウェー
Swiss National Science Foundation
P500PB_203053
スイス
引用
ジャーナル: Nat Struct Mol Biol / 年: 2024 タイトル: Structural basis for excitatory neuropeptide signaling. 著者: Valeria Kalienkova / Mowgli Dandamudi / Cristina Paulino / Timothy Lynagh / 要旨: Rapid signaling between neurons is mediated by ligand-gated ion channels, cell-surface proteins with an extracellular ligand-binding domain and a membrane-spanning ion channel domain. The ...Rapid signaling between neurons is mediated by ligand-gated ion channels, cell-surface proteins with an extracellular ligand-binding domain and a membrane-spanning ion channel domain. The degenerin/epithelial sodium channel (DEG/ENaC) superfamily is diverse in terms of its gating stimuli, with some DEG/ENaCs gated by neuropeptides, and others gated by pH, mechanical force or enzymatic activity. The mechanism by which ligands bind to and activate DEG/ENaCs is poorly understood. Here we dissected the structural basis for neuropeptide-gated activity of a neuropeptide-gated DEG/ENaC, FMRFamide-gated sodium channel 1 (FaNaC1) from the annelid worm Malacoceros fuliginosus, using cryo-electron microscopy. Structures of FaNaC1 in the ligand-free resting state and in several ligand-bound states reveal the ligand-binding site and capture the ligand-induced conformational changes of channel gating, which we verified with complementary mutagenesis experiments. Our results illuminate channel gating in DEG/ENaCs and offer a structural template for experimental dissection of channel pharmacology and ion conduction.
濃度: 1.83 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES 詳細: nanodisc-reconstituted Malacoceros FaNaC1 bound to the full agonist (FMRFamide) in presence of a pore blocker diminazene
試料支持
詳細: at 5 mA / グリッドの材料: GOLD / グリッドのサイズ: 300 divisions/in. / グリッドのタイプ: Quantifoil R1.2/1.3
急速凍結
装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE-PROPANE / 湿度: 100 % / 凍結前の試料温度: 288.15 K