+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-9101 | |||||||||
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タイトル | The 20S supercomplex engaging the SNAP-25 N-terminus (class 2) | |||||||||
マップデータ | The unsharpened map. | |||||||||
試料 |
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キーワード | SNARE / NSF / SNAP / ATPase / AAA / disassembly / synapse / membrane fusion / exocytosis / HYDROLASE | |||||||||
機能・相同性 | 機能・相同性情報 soluble NSF attachment protein activity / Intra-Golgi traffic / Retrograde transport at the Trans-Golgi-Network / COPI-dependent Golgi-to-ER retrograde traffic / COPI-mediated anterograde transport / trans-Golgi Network Vesicle Budding / BLOC-1 complex / SNARE complex disassembly / regulation of delayed rectifier potassium channel activity / myosin head/neck binding ...soluble NSF attachment protein activity / Intra-Golgi traffic / Retrograde transport at the Trans-Golgi-Network / COPI-dependent Golgi-to-ER retrograde traffic / COPI-mediated anterograde transport / trans-Golgi Network Vesicle Budding / BLOC-1 complex / SNARE complex disassembly / regulation of delayed rectifier potassium channel activity / myosin head/neck binding / exocytic insertion of neurotransmitter receptor to postsynaptic membrane / Other interleukin signaling / ribbon synapse / synaptobrevin 2-SNAP-25-syntaxin-1a-complexin II complex / synaptobrevin 2-SNAP-25-syntaxin-1a complex / extrinsic component of presynaptic membrane / Lysosome Vesicle Biogenesis / synaptic vesicle fusion to presynaptic active zone membrane / synaptobrevin 2-SNAP-25-syntaxin-1a-complexin I complex / positive regulation of norepinephrine secretion / Glutamate Neurotransmitter Release Cycle / Norepinephrine Neurotransmitter Release Cycle / COPII-mediated vesicle transport / positive regulation of catecholamine secretion / Acetylcholine Neurotransmitter Release Cycle / Serotonin Neurotransmitter Release Cycle / GABA synthesis, release, reuptake and degradation / protein-containing complex disassembly / Dopamine Neurotransmitter Release Cycle / Golgi Associated Vesicle Biogenesis / zymogen granule membrane / regulated exocytosis / presynaptic dense core vesicle exocytosis / synaptic vesicle docking / regulation of synaptic vesicle priming / storage vacuole / regulation of establishment of protein localization / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / response to gravity / vesicle-mediated transport in synapse / positive regulation of calcium ion-dependent exocytosis / calcium ion-regulated exocytosis of neurotransmitter / vesicle fusion / eosinophil degranulation / vesicle docking / chloride channel inhibitor activity / secretion by cell / ATP-dependent protein disaggregase activity / SNARE complex / SNAP receptor activity / Cargo recognition for clathrin-mediated endocytosis / regulation of exocytosis / regulation of vesicle-mediated transport / Clathrin-mediated endocytosis / LGI-ADAM interactions / calcium-ion regulated exocytosis / hormone secretion / intra-Golgi vesicle-mediated transport / actomyosin / positive regulation of intracellular protein transport / Golgi to plasma membrane protein transport / neurotransmitter secretion / apical protein localization / positive regulation of hormone secretion / Golgi stack / neurotransmitter receptor internalization / protein localization to membrane / ATP-dependent protein binding / neuron projection terminus / positive regulation of ATP-dependent activity / vesicle-fusing ATPase / regulation of synaptic vesicle recycling / insulin secretion / neurotransmitter transport / syntaxin binding / syntaxin-1 binding / clathrin-coated vesicle / SNARE complex assembly / positive regulation of neurotransmitter secretion / Neutrophil degranulation / endosomal transport / synaptic vesicle priming / regulation of synapse assembly / postsynaptic cytosol / myosin binding / positive regulation of receptor recycling / regulation of neuron projection development / positive regulation of exocytosis / modulation of excitatory postsynaptic potential / exocytosis / associative learning / synaptic vesicle exocytosis / voltage-gated potassium channel activity / protein sumoylation / positive regulation of excitatory postsynaptic potential / synaptic vesicle endocytosis / endomembrane system / calcium channel inhibitor activity / long-term memory / response to glucose 類似検索 - 分子機能 | |||||||||
生物種 | Cricetulus griseus (モンゴルキヌゲネズミ) / Rattus norvegicus (ドブネズミ) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.4 Å | |||||||||
データ登録者 | White KI / Zhao M | |||||||||
資金援助 | 米国, 1件
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引用 | ジャーナル: Elife / 年: 2018 タイトル: Structural principles of SNARE complex recognition by the AAA+ protein NSF. 著者: K Ian White / Minglei Zhao / Ucheor B Choi / Richard A Pfuetzner / Axel T Brunger / 要旨: The recycling of SNARE proteins following complex formation and membrane fusion is an essential process in eukaryotic trafficking. A highly conserved AAA+ protein, NSF (-ethylmaleimide sensitive ...The recycling of SNARE proteins following complex formation and membrane fusion is an essential process in eukaryotic trafficking. A highly conserved AAA+ protein, NSF (-ethylmaleimide sensitive factor) and an adaptor protein, SNAP (soluble NSF attachment protein), disassemble the SNARE complex. We report electron-cryomicroscopy structures of the complex of NSF, αSNAP, and the full-length soluble neuronal SNARE complex (composed of syntaxin-1A, synaptobrevin-2, SNAP-25A) in the presence of ATP under non-hydrolyzing conditions at ~3.9 Å resolution. These structures reveal electrostatic interactions by which two αSNAP molecules interface with a specific surface of the SNARE complex. This interaction positions the SNAREs such that the 15 N-terminal residues of SNAP-25A are loaded into the D1 ring pore of NSF via a spiral pattern of interactions between a conserved tyrosine NSF residue and SNAP-25A backbone atoms. This loading process likely precedes ATP hydrolysis. Subsequent ATP hydrolysis then drives complete disassembly. | |||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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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_9101.map.gz | 35.7 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-9101-v30.xml emd-9101.xml | 26 KB 26 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_9101_fsc.xml | 8.1 KB | 表示 | FSCデータファイル |
画像 | emd_9101.png | 48.5 KB | ||
Filedesc metadata | emd-9101.cif.gz | 7.7 KB | ||
その他 | emd_9101_additional.map.gz | 43.5 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-9101 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-9101 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_9101_validation.pdf.gz | 497.5 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_9101_full_validation.pdf.gz | 497.1 KB | 表示 | |
XML形式データ | emd_9101_validation.xml.gz | 10.3 KB | 表示 | |
CIF形式データ | emd_9101_validation.cif.gz | 13.3 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9101 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9101 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「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_9101.map.gz / 形式: CCP4 / 大きさ: 46.4 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | The unsharpened map. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.31 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
-追加マップ: The sharpened map.
ファイル | emd_9101_additional.map | ||||||||||||
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注釈 | The sharpened map. | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
+全体 : 20S supercomplex consisting of soluble neuronal SNARE complex, al...
+超分子 #1: 20S supercomplex consisting of soluble neuronal SNARE complex, al...
+超分子 #2: N-ethylmaleimide sensitive factor
+超分子 #3: Synaptosomal-associated protein 25
+超分子 #4: Syntaxin-1A
+超分子 #5: Vesicle-associated membrane protein 2
+超分子 #6: Alpha-soluble NSF attachment protein
+分子 #1: Vesicle-fusing ATPase
+分子 #2: Synaptosomal-associated protein 25
+分子 #3: Syntaxin-1A
+分子 #4: Vesicle-associated membrane protein 2
+分子 #5: Alpha-soluble NSF attachment protein
+分子 #6: ADENOSINE-5'-TRIPHOSPHATE
+分子 #7: ADENOSINE-5'-DIPHOSPHATE
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
濃度 | 15 mg/mL | |||||||||||||||||||||
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緩衝液 | pH: 8 構成要素:
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グリッド | モデル: Quantifoil R1.2/1.3 / 材質: COPPER / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY ARRAY | |||||||||||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 293 K / 装置: FEI VITROBOT MARK I / 詳細: Blot for 3.5 seconds before plunging.. |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: SUPER-RESOLUTION / デジタル化 - 画像ごとのフレーム数: 2-40 / 撮影したグリッド数: 2 / 実像数: 5418 / 平均露光時間: 10.0 sec. / 平均電子線量: 58.0 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: 70.0 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): 3.0 µm / 最小 デフォーカス(公称値): 1.5 µm |
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 |