ジャーナル: Nat Commun / 年: 2022 タイトル: Structural insights into inhibitory mechanism of human excitatory amino acid transporter EAAT2. 著者: Takafumi Kato / Tsukasa Kusakizako / Chunhuan Jin / Xinyu Zhou / Ryuichi Ohgaki / LiLi Quan / Minhui Xu / Suguru Okuda / Kan Kobayashi / Keitaro Yamashita / Tomohiro Nishizawa / Yoshikatsu ...著者: Takafumi Kato / Tsukasa Kusakizako / Chunhuan Jin / Xinyu Zhou / Ryuichi Ohgaki / LiLi Quan / Minhui Xu / Suguru Okuda / Kan Kobayashi / Keitaro Yamashita / Tomohiro Nishizawa / Yoshikatsu Kanai / Osamu Nureki / 要旨: Glutamate is a pivotal excitatory neurotransmitter in mammalian brains, but excessive glutamate causes numerous neural disorders. Almost all extracellular glutamate is retrieved by the glial ...Glutamate is a pivotal excitatory neurotransmitter in mammalian brains, but excessive glutamate causes numerous neural disorders. Almost all extracellular glutamate is retrieved by the glial transporter, Excitatory Amino Acid Transporter 2 (EAAT2), belonging to the SLC1A family. However, in some cancers, EAAT2 expression is enhanced and causes resistance to therapies by metabolic disturbance. Despite its crucial roles, the detailed structural information about EAAT2 has not been available. Here, we report cryo-EM structures of human EAAT2 in substrate-free and selective inhibitor WAY213613-bound states at 3.2 Å and 2.8 Å, respectively. EAAT2 forms a trimer, with each protomer consisting of transport and scaffold domains. Along with a glutamate-binding site, the transport domain possesses a cavity that could be disrupted during the transport cycle. WAY213613 occupies both the glutamate-binding site and cavity of EAAT2 to interfere with its alternating access, where the sensitivity is defined by the inner environment of the cavity. We provide the characterization of the molecular features of EAAT2 and its selective inhibition mechanism that may facilitate structure-based drug design for EAAT2.
電子線照射量: 50 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k)
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解析
ソフトウェア
名称: REFMAC / バージョン: 5.8.0291 / 分類: 精密化
EMソフトウェア
ID
名称
バージョン
カテゴリ
2
SerialEM
画像取得
7
MOLREP
モデルフィッティング
9
REFMAC
5.8.0291
モデル精密化
13
RELION
3.1
3次元再構成
CTF補正
タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION
対称性
点対称性: C3 (3回回転対称)
3次元再構成
解像度: 3.49 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 527996 / 対称性のタイプ: POINT
精密化
解像度: 3.49→3.49 Å / Cor.coef. Fo:Fc: 0.848 / WRfactor Rwork: 0.345 / SU B: 9.696 / SU ML: 0.184 / Average fsc overall: 0.8451 / Average fsc work: 0.8451 / ESU R: 0.176 / 詳細: Hydrogens have been added in their riding positions /