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データを開く
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基本情報
登録情報 | データベース: EMDB / ID: EMD-3282 | |||||||||
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タイトル | Negative stain electron microscopy structure of TssA from EAEC type VI secretion system | |||||||||
![]() | 3D Reconstruction of TssA dodecamer | |||||||||
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![]() | Type VI protein secretion system / bacterial contractile tail structure | |||||||||
生物種 | ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / ネガティブ染色法 / 解像度: 19.0 Å | |||||||||
![]() | Durand E / Fronzes R / Cambillau C / Cascales E | |||||||||
![]() | ![]() タイトル: Priming and polymerization of a bacterial contractile tail structure. 著者: Abdelrahim Zoued / Eric Durand / Yannick R Brunet / Silvia Spinelli / Badreddine Douzi / Mathilde Guzzo / Nicolas Flaugnatti / Pierre Legrand / Laure Journet / Rémi Fronzes / Tâm Mignot / ...著者: Abdelrahim Zoued / Eric Durand / Yannick R Brunet / Silvia Spinelli / Badreddine Douzi / Mathilde Guzzo / Nicolas Flaugnatti / Pierre Legrand / Laure Journet / Rémi Fronzes / Tâm Mignot / Christian Cambillau / Eric Cascales / ![]() 要旨: Contractile tails are composed of an inner tube wrapped by an outer sheath assembled in an extended, metastable conformation that stores mechanical energy necessary for its contraction. Contraction ...Contractile tails are composed of an inner tube wrapped by an outer sheath assembled in an extended, metastable conformation that stores mechanical energy necessary for its contraction. Contraction is used to propel the rigid inner tube towards target cells for DNA or toxin delivery. Although recent studies have revealed the structure of the contractile sheath of the type VI secretion system, the mechanisms by which its polymerization is controlled and coordinated with the assembly of the inner tube remain unknown. Here we show that the starfish-like TssA dodecameric complex interacts with tube and sheath components. Fluorescence microscopy experiments in enteroaggregative Escherichia coli reveal that TssA binds first to the type VI secretion system membrane core complex and then initiates tail polymerization. TssA remains at the tip of the growing structure and incorporates new tube and sheath blocks. On the basis of these results, we propose that TssA primes and coordinates tail tube and sheath biogenesis. | |||||||||
履歴 |
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構造の表示
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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マップデータ | ![]() | 5.4 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 10.9 KB 10.9 KB | 表示 表示 | ![]() |
画像 | ![]() | 226.4 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 199.7 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 198.8 KB | 表示 | |
XML形式データ | ![]() | 5.4 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
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リンク
EMDBのページ | ![]() ![]() |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | 3D Reconstruction of TssA dodecamer | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.9 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
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試料の構成要素
-全体 : TssA dodecamer
全体 | 名称: TssA dodecamer |
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要素 |
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-超分子 #1000: TssA dodecamer
超分子 | 名称: TssA dodecamer / タイプ: sample / ID: 1000 / 詳細: The sample was mono disperse / 集合状態: 12 / Number unique components: 1 |
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分子量 | 実験値: 952 MDa / 理論値: 891 MDa 手法: on-line multi-angle laser light scattering/quasi-elastic light scattering/absorbance/refractive index (MALS/QELS/UV/RI) |
-分子 #1: TssA
分子 | 名称: TssA / タイプ: protein_or_peptide / ID: 1 / Name.synonym: TssA / コピー数: 12 / 集合状態: Dodecamer / 組換発現: Yes |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 実験値: 74 KDa / 理論値: 74 KDa |
組換発現 | 生物種: ![]() ![]() |
-実験情報
-構造解析
手法 | ネガティブ染色法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
濃度 | 0.01 mg/mL |
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緩衝液 | pH: 8 / 詳細: 20mM Tris-HCL, 150mM NaCl |
染色 | タイプ: NEGATIVE 詳細: TssA sample was applied to glow-discharged carbon-coated copper grids (Agar Scientific). After 30 sec of absorption, the sample was blotted, washed with three drops of water and then stained ...詳細: TssA sample was applied to glow-discharged carbon-coated copper grids (Agar Scientific). After 30 sec of absorption, the sample was blotted, washed with three drops of water and then stained with 2% uranyl acetate |
グリッド | 詳細: 400 mesh glow-discharged carbon-coated copper grids (Agar Scientific) |
凍結 | 凍結剤: NONE / 装置: OTHER |
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電子顕微鏡法
顕微鏡 | FEI TECNAI F20 |
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アライメント法 | Legacy - 非点収差: Objective lens astigmatism was corrected at 100,000 times magnification |
特殊光学系 | エネルギーフィルター - 名称: FEI |
日付 | 2015年6月10日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: FEI FALCON II (4k x 4k) 実像数: 500 / 平均電子線量: 14 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: ![]() |
電子光学系 | 照射モード: SPOT SCAN / 撮影モード: BRIGHT FIELD / Cs: 2.10 mm / 最大 デフォーカス(公称値): 2.5 µm / 最小 デフォーカス(公称値): 0.5 µm / 倍率(公称値): 50000 |
試料ステージ | 試料ホルダーモデル: OTHER |
実験機器 | ![]() モデル: Tecnai F20 / 画像提供: FEI Company |
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画像解析
詳細 | A total of 100000 particles were automatically selected from 500 independent images and extracted within boxes of 180 pixels by 180 pixels using EMAN2/BOXER. The CTF was estimated and corrected by phase flipping using EMAN2. All two- and three-dimensional classifications and refinements were performed using RELION 1.3. We used three rounds of reference-free 2D class averaging to clean up the automatically selected dataset. Only highly populated classes displaying high-resolution features were conserved during this procedure and a final dataset of 20000 particles was assembled. An initial 3D-model was generated in EMAN2 using using 30 classes. 3D classification was then performed in Relion with 5 classes. The particles corresponding to most populated class were used for refinement. Relion auto-refine procedure was used to obtain a final reconstruction at 19A resolution after masking and with D6 symmetry imposed. Reported resolutions are based on the gold-standard Fourier shell correlation 0.143 criterion, and FSC curve were corrected for the effects of a soft mask on the FSC curve using high-resolution noise substitution. |
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CTF補正 | 詳細: Each particle |
最終 再構成 | 想定した対称性 - 点群: D6 (2回x6回 2面回転対称) アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 19.0 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: EMAN2, RELION / 使用した粒子像数: 18000 |
最終 2次元分類 | クラス数: 10 |