+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-22024 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
タイトル | hEAAT3-Asymmetric-2o1i-in20mM-Asp | |||||||||
マップデータ | hEAAT3 asymmetric trimer with 2 outward-facing protomers and 1 inward-facing protomer in 20mM Asp | |||||||||
試料 |
| |||||||||
機能・相同性 | 機能・相同性情報 response to decreased oxygen levels / D-aspartate transmembrane transport / response to anesthetic / regulation of protein targeting to membrane / neurotransmitter receptor transport to plasma membrane / D-aspartate transmembrane transporter activity / Defective SLC1A1 is implicated in schizophrenia 18 (SCZD18) and dicarboxylic aminoaciduria (DCBXA) / distal dendrite / cysteine transmembrane transporter activity / cysteine transport ...response to decreased oxygen levels / D-aspartate transmembrane transport / response to anesthetic / regulation of protein targeting to membrane / neurotransmitter receptor transport to plasma membrane / D-aspartate transmembrane transporter activity / Defective SLC1A1 is implicated in schizophrenia 18 (SCZD18) and dicarboxylic aminoaciduria (DCBXA) / distal dendrite / cysteine transmembrane transporter activity / cysteine transport / monoatomic anion channel activity / high-affinity L-glutamate transmembrane transporter activity / glutamate:sodium symporter activity / L-glutamate import / cellular response to mercury ion / Transport of inorganic cations/anions and amino acids/oligopeptides / conditioned place preference / zinc ion transmembrane transport / L-glutamate transmembrane transport / retina layer formation / cellular response to bisphenol A / L-glutamate transmembrane transporter activity / L-aspartate transmembrane transport / D-aspartate import across plasma membrane / cellular response to ammonium ion / proximal dendrite / glutathione biosynthetic process / righting reflex / L-aspartate transmembrane transporter activity / grooming behavior / L-aspartate import across plasma membrane / intracellular glutamate homeostasis / Glutamate Neurotransmitter Release Cycle / L-glutamate import across plasma membrane / apical dendrite / transepithelial transport / response to morphine / intracellular zinc ion homeostasis / 神経伝達物質輸送体 / glutamate receptor signaling pathway / cellular response to cocaine / chloride transmembrane transporter activity / blood vessel morphogenesis / motor neuron apoptotic process / motor behavior / glutamate binding / dopamine receptor signaling pathway / adult behavior / maintenance of blood-brain barrier / positive regulation of heart rate / dopamine metabolic process / postsynaptic modulation of chemical synaptic transmission / superoxide metabolic process / heart contraction / perisynaptic space / glial cell projection / transport across blood-brain barrier / cellular response to organic cyclic compound / asymmetric synapse / response to axon injury / behavioral fear response / axon terminus / monoatomic ion transport / synaptic cleft / chloride transmembrane transport / response to amphetamine / 神経発生 / dendritic shaft / locomotory behavior / long-term synaptic potentiation / cell periphery / synapse organization / regulation of protein phosphorylation / brain development / Schaffer collateral - CA1 synapse / 記憶 / cytokine-mediated signaling pathway / recycling endosome membrane / presynapse / cellular response to oxidative stress / late endosome membrane / 遺伝子発現 / perikaryon / chemical synaptic transmission / early endosome membrane / negative regulation of neuron apoptotic process / 樹状突起スパイン / response to xenobiotic stimulus / 脂質ラフト / apical plasma membrane / 神経繊維 / neuronal cell body / 樹状突起 / 細胞膜 / 小胞体 / extracellular exosome / 生体膜 / identical protein binding / metal ion binding / 細胞膜 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.71 Å | |||||||||
データ登録者 | Qiu B / Matthies D / Boudker O | |||||||||
資金援助 | 米国, 1件
| |||||||||
引用 | ジャーナル: Sci Adv / 年: 2021 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 |
ムービービューア |
---|---|
構造ビューア | EMマップ: SurfViewMolmilJmol/JSmol |
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 | emd_22024.map.gz | 37.4 MB | EMDBマップデータ形式 | |
---|---|---|---|---|
ヘッダ (付随情報) | emd-22024-v30.xml emd-22024.xml | 12.8 KB 12.8 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_22024_fsc.xml | 7.9 KB | 表示 | FSCデータファイル |
画像 | emd_22024.png | 40.5 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-22024 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-22024 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
---|---|
「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_22024.map.gz / 形式: CCP4 / 大きさ: 40.6 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | hEAAT3 asymmetric trimer with 2 outward-facing protomers and 1 inward-facing protomer in 20mM Asp | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.096 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 : human Excitatory amino acid transporter 3 in 20mM Asp
全体 | 名称: human Excitatory amino acid transporter 3 in 20mM Asp |
---|---|
要素 |
|
-超分子 #1: human Excitatory amino acid transporter 3 in 20mM Asp
超分子 | 名称: human Excitatory amino acid transporter 3 in 20mM Asp タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: all |
---|---|
由来(天然) | 生物種: Homo sapiens (ヒト) |
組換発現 | 生物種: Homo sapiens (ヒト) |
-分子 #1: human Excitatory amino acid transporter 3
分子 | 名称: human 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 光学異性体: LEVO |
---|---|
由来(天然) | 生物種: Homo sapiens (ヒト) |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: GPMGKPARKG CEWKRFLKNN WVLLSTVAAV VLGITTGVLV REHSNLSTLE KFYFAFPGEI LMRMLKLIIL PL IISSMIT GVAALDSNVS GKIGLRAVVY YFCTTLIAVI LGIVLVVSIK PGVTQKVGEI ARTGSTPEVS TVD AMLDLI RNMFPENLVQ ACFQQYKTKR ...文字列: GPMGKPARKG CEWKRFLKNN WVLLSTVAAV VLGITTGVLV REHSNLSTLE KFYFAFPGEI LMRMLKLIIL PL IISSMIT GVAALDSNVS GKIGLRAVVY YFCTTLIAVI LGIVLVVSIK PGVTQKVGEI ARTGSTPEVS TVD AMLDLI RNMFPENLVQ ACFQQYKTKR EEVKPPSDPE MNMTEESFTA VMTTAISKNK TKEYKIVGMY SDGI NVLGL IVFCLVFGLV IGKMGEKGQI LVDFFNALSD ATMKIVQIIM CYMPLGILFL IAGKIIEVED WEIFR KLGL YMATVLTGLA IHSIVILPLI YFIVVRKNPF RFAMGMAQAL LTALMISSSS ATLPVTFRCA EENNQV DKR ITRFVLPVGA TINMDGTALY EAVAAVFIAQ LNDLDLGIGQ IITISITATS ASIGAAGVPQ AGLVTMV IV LSAVGLPAED VTLIIAVDWL LDRFRTMVNV LGDAFGTGIV EKLSKKELEQ MDVSSEVNIV NPFALEST I LDNEDSDTKK SYVNGGFAVD KSDTISFTQT SQF |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
---|---|
解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
濃度 | 5 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 / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 雰囲気: AIR | ||||||||||||||||||
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 |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
---|---|
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: 100.0 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELDBright-field microscopy / Cs: 0.001 mm / 最大 デフォーカス(公称値): 2.0 µm / 最小 デフォーカス(公称値): 1.0 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 |
撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: COUNTING / 平均露光時間: 10.0 sec. / 平均電子線量: 67.6 e/Å2 |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |