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基本情報
登録情報 | データベース: PDB / ID: 8enc | ||||||
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タイトル | Helical reconstruction of the human cardiac actin-tropomyosin-myosin loop 4 7G mutant complex | ||||||
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![]() | MOTOR PROTEIN / actin / tropomyosin / myosin / cardiac | ||||||
機能・相同性 | ![]() positive regulation of heart rate by epinephrine / muscle thin filament tropomyosin / regulation of slow-twitch skeletal muscle fiber contraction / regulation of the force of skeletal muscle contraction / actin-myosin filament sliding / bleb / negative regulation of vascular associated smooth muscle cell migration / regulation of muscle contraction / muscle myosin complex / muscle filament sliding ...positive regulation of heart rate by epinephrine / muscle thin filament tropomyosin / regulation of slow-twitch skeletal muscle fiber contraction / regulation of the force of skeletal muscle contraction / actin-myosin filament sliding / bleb / negative regulation of vascular associated smooth muscle cell migration / regulation of muscle contraction / muscle myosin complex / muscle filament sliding / transition between fast and slow fiber / regulation of the force of heart contraction / myosin filament / ruffle organization / adult heart development / cardiac muscle hypertrophy in response to stress / positive regulation of ATP-dependent activity / Striated Muscle Contraction / myosin complex / ventricular cardiac muscle tissue morphogenesis / structural constituent of muscle / regulation of heart contraction / myosin II complex / microfilament motor activity / myosin binding / myofibril / sarcomere organization / heart contraction / mesenchyme migration / negative regulation of vascular associated smooth muscle cell proliferation / skeletal muscle contraction / positive regulation of cell adhesion / Smooth Muscle Contraction / striated muscle contraction / cardiac muscle contraction / stress fiber / ATP metabolic process / positive regulation of stress fiber assembly / cytoskeleton organization / cytoskeletal protein binding / regulation of heart rate / sarcomere / negative regulation of cell migration / filopodium / muscle contraction / actin filament / actin filament organization / wound healing / structural constituent of cytoskeleton / ruffle membrane / Z disc / cellular response to reactive oxygen species / actin filament binding / actin cytoskeleton / lamellipodium / actin binding / regulation of cell shape / cell body / cytoskeleton / calmodulin binding / protein heterodimerization activity / positive regulation of gene expression / protein homodimerization activity / ATP binding / identical protein binding / cytoplasm / cytosol 類似検索 - 分子機能 | ||||||
生物種 | ![]() ![]() ![]() | ||||||
手法 | 電子顕微鏡法 / らせん対称体再構成法 / クライオ電子顕微鏡法 / 解像度: 3.6 Å | ||||||
![]() | Doran, M.H. / Lehman, W. / Rynkiewicz, M.J. | ||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Myosin loop-4 is critical for optimal tropomyosin repositioning on actin during muscle activation and relaxation. 著者: Matthew H Doran / Michael J Rynkiewicz / Elumalai Pavadai / Skylar M L Bodt / David Rasicci / Jeffrey R Moore / Christopher M Yengo / Esther Bullitt / William Lehman / ![]() 要旨: During force-generating steps of the muscle crossbridge cycle, the tip of the myosin motor, specifically loop-4, contacts the tropomyosin cable of actin filaments. In the current study, we determined ...During force-generating steps of the muscle crossbridge cycle, the tip of the myosin motor, specifically loop-4, contacts the tropomyosin cable of actin filaments. In the current study, we determined the corresponding effect of myosin loop-4 on the regulatory positioning of tropomyosin on actin. To accomplish this, we compared high-resolution cryo-EM structures of myosin S1-decorated thin filaments containing either wild-type or a loop-4 mutant construct, where the seven-residue portion of myosin loop-4 that contacts tropomyosin was replaced by glycine residues, thus removing polar side chains from residues 366-372. Cryo-EM analysis of fully decorated actin-tropomyosin filaments with wild-type and mutant S1, yielded 3.4-3.6 Å resolution reconstructions, with even higher definition at the actin-myosin interface. Loop-4 densities both in wild-type and mutant S1 were clearly identified, and side chains were resolved in the wild-type structure. Aside from loop-4, actin and myosin structural domains were indistinguishable from each other when filaments were decorated with either mutant or wild-type S1. In marked contrast, the position of tropomyosin on actin in the two reconstructions differed by 3 to 4 Å. In maps of filaments containing the mutant, tropomyosin was located closer to the myosin-head and thus moved in the direction of the C-state conformation adopted by myosin-free thin filaments. Complementary interaction energy measurements showed that tropomyosin in the mutant thin filaments sits on actin in a local energy minimum, whereas tropomyosin is positioned by wild-type S1 in an energetically unfavorable location. We propose that the high potential energy associated with tropomyosin positioning in wild-type filaments favors an effective transition to B- and C-states following release of myosin from the thin filaments during relaxation. | ||||||
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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PDBx/mmCIF形式 | ![]() | 533.9 KB | 表示 | ![]() |
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PDB形式 | ![]() | 422.9 KB | 表示 | ![]() |
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-検証レポート
文書・要旨 | ![]() | 1.5 MB | 表示 | ![]() |
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文書・詳細版 | ![]() | 1.5 MB | 表示 | |
XML形式データ | ![]() | 92.7 KB | 表示 | |
CIF形式データ | ![]() | 139 KB | 表示 | |
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 | ![]() 28270MC ![]() 8efiC M: このデータのモデリングに利用したマップデータ C: 同じ文献を引用 ( |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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リンク
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集合体
登録構造単位 | ![]()
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要素
#1: タンパク質 | 分子量: 222931.328 Da / 分子数: 1 / 変異: Residues 366-372 substituted with seven glycines / 由来タイプ: 組換発現 / 由来: (組換発現) ![]() ![]() ![]() | ||||||||
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#2: タンパク質 | 分子量: 42064.891 Da / 分子数: 5 / 由来タイプ: 天然 / 由来: (天然) ![]() ![]() #3: タンパク質 | 分子量: 32763.621 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) ![]() ![]() ![]() #4: 化合物 | ChemComp-ADP / #5: 化合物 | ChemComp-MG / 研究の焦点であるリガンドがあるか | N | |
-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: FILAMENT / 3次元再構成法: らせん対称体再構成法 |
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試料調製
構成要素 |
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緩衝液 | pH: 7 | |||||||||||||||||||||||||||||||||||
試料 | 濃度: 0.13 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | |||||||||||||||||||||||||||||||||||
試料支持 | グリッドの材料: GOLD / グリッドのサイズ: 400 divisions/in. / グリッドのタイプ: Quantifoil R1.2/1.3 | |||||||||||||||||||||||||||||||||||
急速凍結 | 装置: FEI VITROBOT MARK III / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 283 K |
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電子顕微鏡撮影
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: TFS KRIOS |
電子銃 | 電子線源: ![]() |
電子レンズ | モード: BRIGHT FIELD / 倍率(公称値): 80000 X / 最大 デフォーカス(公称値): 3000 nm / 最小 デフォーカス(公称値): 700 nm / Cs: 2.7 mm |
試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | 平均露光時間: 3.12 sec. / 電子線照射量: 54 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 撮影したグリッド数: 4 / 実像数: 4171 |
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解析
ソフトウェア | 名称: PHENIX / バージョン: 1.18_3861: / 分類: 精密化 | ||||||||||||||||||||||||||||||||||||
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EMソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING ONLY | ||||||||||||||||||||||||||||||||||||
らせん対称 | 回転角度/サブユニット: 166.8 ° / 軸方向距離/サブユニット: 27.9 Å / らせん対称軸の対称性: C1 | ||||||||||||||||||||||||||||||||||||
粒子像の選択 | 選択した粒子像数: 655287 | ||||||||||||||||||||||||||||||||||||
3次元再構成 | 解像度: 3.6 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 187885 / 対称性のタイプ: HELICAL | ||||||||||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: OTHER / 空間: REAL | ||||||||||||||||||||||||||||||||||||
原子モデル構築 | PDB-ID: 6X5Z Accession code: 6X5Z / Source name: PDB / タイプ: experimental model | ||||||||||||||||||||||||||||||||||||
拘束条件 |
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