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基本情報
登録情報 | データベース: EMDB / ID: EMD-1585 | |||||||||
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タイトル | Electron tomography of isometrically contracting insect flight muscle quick frozen after a rapid stretch transient | |||||||||
![]() | This is a global average of aligned subvolumes. | |||||||||
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![]() | insect / muscle / myosin / troponin / tropomyosin / actin / light chains / thin filament / thick filament / electron microscopy / image processing / isometric contraction / freezing / freeze substitution / microtomy / multivariate data analysis | |||||||||
機能・相同性 | ![]() cytoskeletal motor regulator activity / mitotic actomyosin contractile ring / troponin C binding / mitotic actomyosin contractile ring contraction / contractile muscle fiber / troponin complex / Striated Muscle Contraction / regulation of muscle contraction / muscle myosin complex / myosin filament ...cytoskeletal motor regulator activity / mitotic actomyosin contractile ring / troponin C binding / mitotic actomyosin contractile ring contraction / contractile muscle fiber / troponin complex / Striated Muscle Contraction / regulation of muscle contraction / muscle myosin complex / myosin filament / actomyosin structure organization / myosin II complex / locomotion / myosin complex / cytoskeletal motor activator activity / sarcomere organization / microfilament motor activity / myofibril / tropomyosin binding / mesenchyme migration / troponin I binding / myosin heavy chain binding / filamentous actin / actin filament bundle / skeletal muscle thin filament assembly / striated muscle thin filament / actin filament bundle assembly / skeletal muscle contraction / mitotic cytokinesis / skeletal muscle myofibril / actin monomer binding / skeletal muscle fiber development / muscle contraction / cardiac muscle contraction / stress fiber / skeletal muscle tissue development / titin binding / actin filament polymerization / post-embryonic development / filopodium / actin filament organization / actin filament / 加水分解酵素; 酸無水物に作用; 酸無水物に作用・細胞または細胞小器官の運動に関与 / calcium-dependent protein binding / actin filament binding / actin cytoskeleton / lamellipodium / actin binding / cell body / calmodulin binding / hydrolase activity / protein heterodimerization activity / protein domain specific binding / calcium ion binding / positive regulation of gene expression / magnesium ion binding / protein homodimerization activity / ATP binding / identical protein binding / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | ![]() | |||||||||
手法 | サブトモグラム平均法 / クライオ電子顕微鏡法 / ネガティブ染色法 | |||||||||
![]() | 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. | |||||||||
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構造ビューア | EMマップ: ![]() ![]() ![]() |
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マップデータ | ![]() | 1.1 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 13 KB 13 KB | 表示 表示 | ![]() |
画像 | ![]() | 86.2 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 | ![]() 2w4gM ![]() 2w4hMC ![]() 2w4uMC ![]() 2w4vMC ![]() 2w4wMC ![]() 1584C C: 同じ文献を引用 ( M: このマップから作成された原子モデル |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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注釈 | This is a global average of aligned subvolumes. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 6.9 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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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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試料の構成要素
-全体 : Isometrically contracting asynchronous insect flight muscle upon ...
全体 | 名称: Isometrically contracting asynchronous insect flight muscle upon a quick stretch |
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要素 |
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-超分子 #1000: Isometrically contracting asynchronous insect flight muscle upon ...
超分子 | 名称: Isometrically contracting asynchronous insect flight muscle upon a quick stretch 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. The fiber ...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. The fiber was stretched 6 nm per half-sarcomere in 2 ms and length was held constant after the length step. The freezing impact occurred 6-7 ms later. 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 | ネガティブ染色法, クライオ電子顕微鏡法 |
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![]() | サブトモグラム平均法 |
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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 Timed resolved state: Frozen 6-7 msec after application of the length transient 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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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 |
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 / Cs: 2.0 mm |
試料ステージ | 試料ホルダー: side entry, eucentric / 試料ホルダーモデル: OTHER / Tilt series - Axis1 - Min angle: -72 ° / Tilt series - Axis1 - Max angle: 72 ° |
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画像解析
詳細 | The fiber was stretched 6 nm per half-sarcomere in 2 ms and length was held constant after the length step. The freezing impact occurred 6-7 ms later. Average number of tilts used in the 3D reconstructions: 70. |
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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. |