+
データを開く
-
基本情報
登録情報 | データベース: EMDB / ID: EMD-22021 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
タイトル | hEAAT3-IFS-Apo | |||||||||
![]() | hEAAT3 inward-facing Apo state | |||||||||
![]() |
| |||||||||
![]() | hEAAT3 inward-facing Apo state / TRANSPORT PROTEIN | |||||||||
機能・相同性 | ![]() D-aspartate transmembrane transport / regulation of protein targeting to membrane / D-aspartate transmembrane transporter activity / Defective SLC1A1 is implicated in schizophrenia 18 (SCZD18) and dicarboxylic aminoaciduria (DCBXA) / distal dendrite / cysteine transport / cysteine transmembrane transporter activity / neurotransmitter receptor transport to plasma membrane / high-affinity L-glutamate transmembrane transporter activity / glutamate:sodium symporter activity ...D-aspartate transmembrane transport / regulation of protein targeting to membrane / D-aspartate transmembrane transporter activity / Defective SLC1A1 is implicated in schizophrenia 18 (SCZD18) and dicarboxylic aminoaciduria (DCBXA) / distal dendrite / cysteine transport / cysteine transmembrane transporter activity / neurotransmitter receptor transport to plasma membrane / high-affinity L-glutamate transmembrane transporter activity / glutamate:sodium symporter activity / response to decreased oxygen levels / L-glutamate import / cellular response to mercury ion / Transport of inorganic cations/anions and amino acids/oligopeptides / retina layer formation / L-glutamate transmembrane transporter activity / L-glutamate transmembrane transport / glutathione biosynthetic process / D-aspartate import across plasma membrane / L-aspartate transmembrane transport / cellular response to ammonium ion / righting reflex / zinc ion transmembrane transport / cellular response to bisphenol A / L-aspartate transmembrane transporter activity / grooming behavior / intracellular glutamate homeostasis / L-aspartate import across plasma membrane / Glutamate Neurotransmitter Release Cycle / monoatomic anion channel activity / L-glutamate import across plasma membrane / proximal dendrite / transepithelial transport / apical dendrite / intracellular zinc ion homeostasis / conditioned place preference / cellular response to cocaine / blood vessel morphogenesis / chloride transmembrane transporter activity / motor neuron apoptotic process / response to morphine / G protein-coupled dopamine receptor signaling pathway / glutamate receptor signaling pathway / response to anesthetic / neurotransmitter transport / maintenance of blood-brain barrier / superoxide metabolic process / heart contraction / perisynaptic space / dopamine metabolic process / motor behavior / adult behavior / asymmetric synapse / glial cell projection / behavioral fear response / postsynaptic modulation of chemical synaptic transmission / synaptic cleft / response to axon injury / positive regulation of heart rate / transport across blood-brain barrier / monoatomic ion transport / axon terminus / neurogenesis / response to amphetamine / chloride transmembrane transport / dendritic shaft / cell periphery / locomotory behavior / synapse organization / brain development / cytokine-mediated signaling pathway / Schaffer collateral - CA1 synapse / long-term synaptic potentiation / memory / recycling endosome membrane / late endosome membrane / presynapse / cellular response to oxidative stress / early endosome membrane / chemical synaptic transmission / dendritic spine / perikaryon / negative regulation of neuron apoptotic process / gene expression / apical plasma membrane / response to xenobiotic stimulus / membrane raft / axon / neuronal cell body / dendrite / cell surface / extracellular exosome / metal ion binding / identical protein binding / membrane / plasma membrane 類似検索 - 分子機能 | |||||||||
生物種 | ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.03 Å | |||||||||
![]() | Qiu B / Matthies D | |||||||||
資金援助 | ![]()
| |||||||||
![]() | ![]() タイトル: Cryo-EM structures of excitatory amino acid transporter 3 visualize coupled substrate, sodium, and proton binding and transport. 著者: Biao Qiu / Doreen Matthies / Eva Fortea / Zhiheng Yu / Olga Boudker / ![]() 要旨: Human excitatory amino acid transporter 3 (hEAAT3) mediates glutamate uptake in neurons, intestine, and kidney. Here, we report cryo-EM structures of hEAAT3 in several functional states where the ...Human excitatory amino acid transporter 3 (hEAAT3) mediates glutamate uptake in neurons, intestine, and kidney. Here, we report cryo-EM structures of hEAAT3 in several functional states where the transporter is empty, bound to coupled sodium ions only, or fully loaded with three sodium ions, a proton, and the substrate aspartate. The structures suggest that hEAAT3 operates by an elevator mechanism involving three functionally independent subunits. When the substrate-binding site is near the cytoplasm, it has a remarkably low affinity for the substrate, perhaps facilitating its release and allowing the rapid transport turnover. The mechanism of the coupled uptake of the sodium ions and the substrate is conserved across evolutionarily distant families and is augmented by coupling to protons in EAATs. The structures further suggest a mechanism by which a conserved glutamate residue mediates proton symport. | |||||||||
履歴 |
|
-
構造の表示
ムービー |
![]() |
---|---|
構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
-
ダウンロードとリンク
-EMDBアーカイブ
マップデータ | ![]() | 84.6 MB | ![]() | |
---|---|---|---|---|
ヘッダ (付随情報) | ![]() ![]() | 15.6 KB 15.6 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 10.3 KB | 表示 | ![]() |
画像 | ![]() | 43.1 KB | ||
Filedesc metadata | ![]() | 6.2 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 553.3 KB | 表示 | ![]() |
---|---|---|---|---|
文書・詳細版 | ![]() | 552.9 KB | 表示 | |
XML形式データ | ![]() | 11.5 KB | 表示 | |
CIF形式データ | ![]() | 15.3 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
-
リンク
EMDBのページ | ![]() ![]() |
---|---|
「今月の分子」の関連する項目 |
-
マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | hEAAT3 inward-facing Apo state | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 0.832 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
|
-添付データ
-
試料の構成要素
-全体 : inward facing hEAAT3 trimer Apo state
全体 | 名称: inward facing hEAAT3 trimer Apo state |
---|---|
要素 |
|
-超分子 #1: inward facing hEAAT3 trimer Apo state
超分子 | 名称: inward facing hEAAT3 trimer Apo state / タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1 |
---|---|
由来(天然) | 生物種: ![]() |
-分子 #1: Excitatory amino acid transporter 3
分子 | 名称: Excitatory amino acid transporter 3 / タイプ: protein_or_peptide / ID: 1 詳細: The Glycine and Proline at the N terminal are the residues left after PreScission Protease treatment コピー数: 3 / 光学異性体: LEVO |
---|---|
由来(天然) | 生物種: ![]() |
分子量 | 理論値: 57.301168 KDa |
組換発現 | 生物種: ![]() |
配列 | 文字列: GPMGKPARKG CEWKRFLKNN WVLLSTVAAV VLGITTGVLV REHSNLSTLE KFYFAFPGEI LMRMLKLIIL PLIISSMITG VAALDSNVS GKIGLRAVVY YFCTTLIAVI LGIVLVVSIK PGVTQKVGEI ARTGSTPEVS TVDAMLDLIR NMFPENLVQA C FQQYKTKR ...文字列: GPMGKPARKG CEWKRFLKNN WVLLSTVAAV VLGITTGVLV REHSNLSTLE KFYFAFPGEI LMRMLKLIIL PLIISSMITG VAALDSNVS GKIGLRAVVY YFCTTLIAVI LGIVLVVSIK PGVTQKVGEI ARTGSTPEVS TVDAMLDLIR NMFPENLVQA C FQQYKTKR EEVKPPSDPE MNMTEESFTA VMTTAISKNK TKEYKIVGMY SDGINVLGLI VFCLVFGLVI GKMGEKGQIL VD FFNALSD ATMKIVQIIM CYMPLGILFL IAGKIIEVED WEIFRKLGLY MATVLTGLAI HSIVILPLIY FIVVRKNPFR FAM GMAQAL LTALMISSSS ATLPVTFRCA EENNQVDKRI TRFVLPVGAT INMDGTALYE AVAAVFIAQL NDLDLGIGQI ITIS ITATS ASIGAAGVPQ AGLVTMVIVL SAVGLPAEDV TLIIAVDWLL DRFRTMVNVL GDAFGTGIVE KLSKKELEQM DVSSE VNIV NPFALESTIL DNEDSDTKKS YVNGGFAVDK SDTISFTQTS QF UniProtKB: Excitatory amino acid transporter 3 |
-分子 #2: CHOLINE ION
分子 | 名称: CHOLINE ION / タイプ: ligand / ID: 2 / コピー数: 3 / 式: CHT |
---|---|
分子量 | 理論値: 104.171 Da |
Chemical component information | ![]() ChemComp-CHT: |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
---|---|
![]() | 単粒子再構成法 |
試料の集合状態 | particle |
-
試料調製
濃度 | 6 mg/mL | |||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
緩衝液 | pH: 8 構成要素:
| |||||||||||||||
グリッド | モデル: Quantifoil R1.2/1.3 / 材質: GOLD / メッシュ: 300 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY | |||||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 K / 装置: FEI VITROBOT MARK IV / 詳細: blot 3s. | |||||||||||||||
詳細 | This sample was mono disperse |
-
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 K3 BIOQUANTUM (6k x 4k) 平均電子線量: 60.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | C2レンズ絞り径: 100.0 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 最大 デフォーカス(公称値): 1.5 µm / 最小 デフォーカス(公称値): 0.5 µm / 倍率(公称値): 105000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |