+
データを開く
-
基本情報
登録情報 | データベース: EMDB / ID: EMD-3301 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
タイトル | The Structure of the Relaxed Thick Filaments from Lethocerus Flight Muscle | |||||||||
![]() | CryoEM IHRSR of relaxed thick filament from Lethocerus indicus (water bug) flight muscle | |||||||||
![]() |
| |||||||||
![]() | thick filament / insect fight muscle / myosin / paramyosin / myofilin / flightin / relaxed state / muscle contraction | |||||||||
機能・相同性 | Myofilin / Myofilin / Myofilin / Flightin / Myofilin protein![]() | |||||||||
生物種 | ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / ネガティブ染色法 / 解像度: 5.5 Å | |||||||||
![]() | Hu Z / Taylor DW / Reedy MK / Edwards RJ / Taylor KA | |||||||||
![]() | ![]() タイトル: Structure of myosin filaments from relaxed flight muscle by cryo-EM at 6 Å resolution. 著者: Zhongjun Hu / Dianne W Taylor / Michael K Reedy / Robert J Edwards / Kenneth A Taylor / ![]() 要旨: We describe a cryo-electron microscopy three-dimensional image reconstruction of relaxed myosin II-containing thick filaments from the flight muscle of the giant water bug . The relaxed thick ...We describe a cryo-electron microscopy three-dimensional image reconstruction of relaxed myosin II-containing thick filaments from the flight muscle of the giant water bug . The relaxed thick filament structure is a key element of muscle physiology because it facilitates the reextension process following contraction. Conversely, the myosin heads must disrupt their relaxed arrangement to drive contraction. Previous models predicted that myosin was unique in having an intermolecular head-head interaction, as opposed to the intramolecular head-head interaction observed in all other species. In contrast to the predicted model, we find an intramolecular head-head interaction, which is similar to that of other thick filaments but oriented in a distinctly different way. The arrangement of myosin's long α-helical coiled-coil rod domain has been hypothesized as either curved layers or helical subfilaments. Our reconstruction is the first report having sufficient resolution to track the rod α helices in their native environment at resolutions ~5.5 Å, and it shows that the layer arrangement is correct for . Threading separate paths through the forest of myosin coiled coils are four nonmyosin peptides. We suggest that the unusual position of the heads and the rod arrangement separated by nonmyosin peptides are adaptations for mechanical signal transduction whereby applied tension disrupts the myosin heads as a component of stretch activation. | |||||||||
履歴 |
|
-
構造の表示
ムービー |
![]() |
---|---|
構造ビューア | EMマップ: ![]() ![]() ![]() |
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 | ![]() | 283.3 MB | ![]() | |
---|---|---|---|---|
ヘッダ (付随情報) | ![]() ![]() | 15.4 KB 15.4 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 15.2 KB | 表示 | ![]() |
画像 | ![]() | 198.3 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 336 KB | 表示 | ![]() |
---|---|---|---|---|
文書・詳細版 | ![]() | 335.1 KB | 表示 | |
XML形式データ | ![]() | 15.1 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
-
リンク
EMDBのページ | ![]() ![]() |
---|
-
マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | CryoEM IHRSR of relaxed thick filament from Lethocerus indicus (water bug) flight muscle | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.223 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 : Thick filament from Lethocerus (waterbug) flight muscle
全体 | 名称: Thick filament from Lethocerus (waterbug) flight muscle |
---|---|
要素 |
|
-超分子 #1000: Thick filament from Lethocerus (waterbug) flight muscle
超分子 | 名称: Thick filament from Lethocerus (waterbug) flight muscle タイプ: sample / ID: 1000 詳細: The sample is a bipolar helical structure, with helical repeat 145 Angstrom and helical turn 33.98 degree. The sample has C4 symmetry. The map contains 6 unique features: myosin molecule with ...詳細: The sample is a bipolar helical structure, with helical repeat 145 Angstrom and helical turn 33.98 degree. The sample has C4 symmetry. The map contains 6 unique features: myosin molecule with completely resolved rods, 4 resolved non-myosin densities among the myosin rods and an annular region inside of annulus occupied by myosin rods that most likely contains paramyosin. The 4 non-myosin densities may contain parts of the proteins myofilin and flightin. 集合状態: bipolar helical structure / Number unique components: 4 |
---|
-分子 #1: myosin II
分子 | 名称: myosin II / タイプ: protein_or_peptide / ID: 1 詳細: The resolution of LMM and C-terminal of S2 domain is about 5.5 Angstrom; the N-terminal (at first crown) of S2 is about 7 angstrom; free head is about 10 angstrom; block head is about 20 angstrom. 集合状態: Helical assembly / 組換発現: No / データベース: NCBI |
---|---|
由来(天然) | 生物種: ![]() |
分子量 | 理論値: 520 KDa |
-分子 #2: paramyosin
分子 | 名称: paramyosin / タイプ: protein_or_peptide / ID: 2 詳細: Paramyosin is located in the filament core, and may not have the same symmetry with myosin. The resolution is lower at 10 Angstrom. 集合状態: dimer / 組換発現: No / データベース: NCBI |
---|---|
由来(天然) | 生物種: ![]() |
分子量 | 理論値: 100 KDa |
-分子 #3: flightin
分子 | 名称: flightin / タイプ: protein_or_peptide / ID: 3 詳細: We can see some non-myosin densities in the reconstruction. Their identification as flightin is uncertain 組換発現: No / データベース: NCBI |
---|---|
由来(天然) | 生物種: ![]() |
分子量 | 理論値: 19 KDa |
配列 | UniProtKB: Flightin |
-分子 #4: myofilin
分子 | 名称: myofilin / タイプ: protein_or_peptide / ID: 4 詳細: We can see some non-myosin densities in the reconstruction. Their identification as myofilin is uncertain. 組換発現: No / データベース: NCBI |
---|---|
由来(天然) | 生物種: ![]() |
分子量 | 理論値: 30.3 KDa |
配列 | UniProtKB: Myofilin protein / InterPro: Myofilin |
-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 |
---|---|
![]() | 単粒子再構成法 |
試料の集合状態 | filament |
-
試料調製
緩衝液 | pH: 6.8 詳細: 8 mM MOPS, 10 mM Na acetate, 1 mM Mg acetate, 1 mM ATP, and 1 mM EGTA |
---|---|
染色 | タイプ: NEGATIVE 詳細: All reconstruction data obtained from unstained, frozen hydrated samples. |
グリッド | 詳細: R2/1 Quantifoil grid |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 100 K / 装置: FEI VITROBOT MARK IV 詳細: The Vitrobot environmental chamber was set to 100% relative humidity for freezing and maintained at a temperature of 22 degrees Celsius. The crude filament prep was applied to the grid, ...詳細: The Vitrobot environmental chamber was set to 100% relative humidity for freezing and maintained at a temperature of 22 degrees Celsius. The crude filament prep was applied to the grid, incubated for 60 seconds, blotted once and then plunged into liquid ethane to vitrify. 手法: EM samples of relaxed filaments were prepared by applying 4 microliters of the above prep to a R2/1 Quantifoil grid at room temperature for 1 minute, washing with several drops of rinse ...手法: EM samples of relaxed filaments were prepared by applying 4 microliters of the above prep to a R2/1 Quantifoil grid at room temperature for 1 minute, washing with several drops of rinse buffer, identical to the relaxing buffer except NaCl was replaced with Na acetate. Following a 2 minute incubation, the grid was washed with several drops of a low salt rinse consisting of 8 mM MOPS, 10 mM Na Acetate, 1 mM Mg Acetate, 1 mM ATP, and 1 mM EGTA, pH 6.8. |
-
電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
---|---|
温度 | 最低: 90 K |
詳細 | 48 frames were recorded over a 1.5 sec exposure time |
日付 | 2015年3月27日 |
撮影 | カテゴリ: CCD / フィルム・検出器のモデル: OTHER / 実像数: 4000 / 平均電子線量: 65 e/Å2 / 詳細: The total dose is 65 electrons with 48 frames / ビット/ピクセル: 32 |
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: 2.7 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): 1.5 µm / 倍率(公称値): 29000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |