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基本情報
登録情報 | データベース: EMDB / ID: EMD-1561 | |||||||||
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タイトル | Electron tomography of isometrically contracting insect flight muscle | |||||||||
![]() | This is a dual axis tomogram of quick frozen, freeze This is a dual axis tomogram of quick frozen, freeze substituted, plastic embedded, thin sectioned insect flight muscle from Lethocerus sp. | |||||||||
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機能・相同性 | ![]() cytoskeletal motor regulator activity / ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() | |||||||||
手法 | ![]() ![]() ![]() | |||||||||
![]() | Wu S / Liu J / Reedy MC / Tregear RT / Winkler H / Franzini-Armstrong C / Sasaki H / Lucaveche C / Goldman YE / Reedy MK / Taylor KA | |||||||||
![]() | ジャーナル: J Struct Biol / 年: 2009 タイトル: Methods for identifying and averaging variable molecular conformations in tomograms of actively contracting insect flight muscle. 著者: Shenping Wu / Jun Liu / Mary C Reedy / Hanspeter Winkler / Michael K Reedy / Kenneth A Taylor / ![]() 要旨: During active muscle contraction, tension is generated through many simultaneous, independent interactions between the molecular motor myosin and the actin filaments. The ensemble of myosin motors ...During active muscle contraction, tension is generated through many simultaneous, independent interactions between the molecular motor myosin and the actin filaments. The ensemble of myosin motors displays heterogeneous conformations reflecting different mechanochemical steps of the ATPase pathway. We used electron tomography of actively contracting insect flight muscle fast-frozen, freeze substituted, Araldite embedded, thin-sectioned and stained, to obtain 3D snapshots of the multiplicity of actin-attached myosin structures. We describe procedures for alignment of the repeating lattice of sub-volumes (38.7 nm cross-bridge repeats bounded by troponin) and multivariate data analysis to identify self-similar repeats for computing class averages. Improvements in alignment and classification of repeat sub-volumes reveals (for the first time in active muscle images) the helix of actin subunits in the thin filament and the troponin density with sufficient clarity that a quasiatomic model of the thin filament can be built into the class averages independent of the myosin cross-bridges. We show how quasiatomic model building can identify both strong and weak myosin attachments to actin. We evaluate the accuracy of image classification to enumerate the different types of actin-myosin attachments. | |||||||||
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: ![]() ![]() ![]() |
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マップデータ | ![]() | 532.4 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 11.3 KB 11.3 KB | 表示 表示 | ![]() |
画像 | ![]() | 189 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 2w49MPC ![]() 2w4aMPC ![]() 2w4tMPC ![]() 2w4gC |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | This is a dual axis tomogram of quick frozen, freeze This is a dual axis tomogram of quick frozen, freeze substituted, plastic embedded, thin sectioned insect flight muscle from Lethocerus sp. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 これらの図は立方格子座標系で作成されたものです | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 6.9 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 : Isometrically contracting asynchronous insect flight muscle
全体 | 名称: Isometrically contracting asynchronous insect flight muscle |
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要素 |
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-超分子 #1000: Isometrically contracting asynchronous insect flight muscle
超分子 | 名称: Isometrically contracting asynchronous insect flight muscle タイプ: sample / ID: 1000 詳細: This specimen is obtained from a quick frozen, isometrically contracting asynchronous insect flight muscle that has been freeze substituted, plastic embedded, and thin sectioned 集合状態: tissue / Number unique components: 8 |
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-超分子 #1: myofibril
超分子 | 名称: myofibril / タイプ: organelle_or_cellular_component / ID: 1 / Name.synonym: muscle 詳細: The sample was enblock stained using uranyl acetate and tannic acid, and post stained with lead citrate and potassium permanganate 集合状態: muscle fibril / 組換発現: No / データベース: NCBI |
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由来(天然) | 生物種: ![]() ![]() |
-実験情報
-構造解析
手法 | ![]() ![]() |
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試料調製
緩衝液 | 詳細: 20 mM MOPS buffer, 5 mM NaN3, and MgCl2, ATP, CaCl2, and EGTA in varying millimolar concentrations |
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染色 | タイプ: NEGATIVE 詳細: Freeze slammed fibers were freeze-substituted in acetone using a tannic aciduranyl acetate sequence, and ultimately embedded in Araldite-506 for thin-section electron microscopy. Ultrathin ...詳細: Freeze slammed fibers were freeze-substituted in acetone using a tannic aciduranyl acetate sequence, and ultimately embedded in Araldite-506 for thin-section electron microscopy. Ultrathin (25-30 nm) longitudinal sections were stained by permanganate-lead. |
凍結 | 凍結剤: HELIUM / チャンバー内温度: 4.5 K / 装置: HOMEMADE PLUNGER 詳細: Vitrification instrument: modified Heuser Cryopress freezing head 手法: smash against a liquid helium cooled Au-coated Cu mirror |
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電子顕微鏡法
顕微鏡 | FEI/PHILIPS CM300FEG/T |
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電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD![]() |
試料ステージ | 試料ホルダー: Gatan model 670 Ultrahigh tilt analytical holder 試料ホルダーモデル: OTHER / Tilt series - Axis1 - Min angle: -72 ° / Tilt series - Axis1 - Max angle: 72 ° |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: TVIPS TEMCAM-F224 (2k x 2k) 詳細: Images recorded on a TVIPS Tem-Cam F224 2k x 2k CCD camera ビット/ピクセル: 16 |
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画像解析
最終 再構成 | アルゴリズム: OTHER / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア - 名称: PROTOMO, IMOD 詳細: Two tilt series at 90 degrees to one another were first independently aligned using marker-free alignment and area matching. The two resulting tomograms were then merged by patch correlation ...詳細: Two tilt series at 90 degrees to one another were first independently aligned using marker-free alignment and area matching. The two resulting tomograms were then merged by patch correlation and volume warp using IMOD. Tomogram computed using weighted back projection. 使用した粒子像数: 70 |
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