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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: PDB / ID: 2dfs | ||||||
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| タイトル | 3-D structure of Myosin-V inhibited state | ||||||
要素 |
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キーワード | CONTRACTILE PROTEIN/TRANSPORT PROTEIN / myosin-V / inhibited state / calmodulin / cryoelectron tomography / CONTRACTILE PROTEIN-TRANSPORT PROTEIN COMPLEX | ||||||
| 機能・相同性 | 機能・相同性情報negative regulation of calcium ion transmembrane transporter activity / CaMK IV-mediated phosphorylation of CREB / Cam-PDE 1 activation / CREB1 phosphorylation through the activation of CaMKII/CaMKK/CaMKIV cascasde / Glycogen breakdown (glycogenolysis) / Activation of RAC1 downstream of NMDARs / Sodium/Calcium exchangers / Activation of Ca-permeable Kainate Receptor / Synthesis of IP3 and IP4 in the cytosol / CLEC7A (Dectin-1) induces NFAT activation ...negative regulation of calcium ion transmembrane transporter activity / CaMK IV-mediated phosphorylation of CREB / Cam-PDE 1 activation / CREB1 phosphorylation through the activation of CaMKII/CaMKK/CaMKIV cascasde / Glycogen breakdown (glycogenolysis) / Activation of RAC1 downstream of NMDARs / Sodium/Calcium exchangers / Activation of Ca-permeable Kainate Receptor / Synthesis of IP3 and IP4 in the cytosol / CLEC7A (Dectin-1) induces NFAT activation / RHO GTPases activate PAKs / Calmodulin induced events / Inactivation, recovery and regulation of the phototransduction cascade / Tetrahydrobiopterin (BH4) synthesis, recycling, salvage and regulation / eNOS activation / Reduction of cytosolic Ca++ levels / Unblocking of NMDA receptors, glutamate binding and activation / Protein methylation / Calcineurin activates NFAT / Ion transport by P-type ATPases / RAF activation / minus-end directed microfilament motor activity / VEGFR2 mediated vascular permeability / RAS processing / Ca2+ pathway / insulin-responsive compartment / RHO GTPases activate IQGAPs / Extra-nuclear estrogen signaling / FCERI mediated Ca+2 mobilization / RAF/MAP kinase cascade / PKA activation / regulation of response to tumor cell / positive regulation of autophagic cell death / DAPK1-calmodulin complex / Smooth Muscle Contraction / : / : / : / : / Platelet degranulation / High laminar flow shear stress activates signaling by PIEZO1 and PECAM1:CDH5:KDR in endothelial cells / : / Stimuli-sensing channels / establishment of protein localization to mitochondrial membrane / Ion homeostasis / type 3 metabotropic glutamate receptor binding / myosin complex / response to corticosterone / negative regulation of high voltage-gated calcium channel activity / negative regulation of ryanodine-sensitive calcium-release channel activity / organelle localization by membrane tethering / microfilament motor activity / mitochondrion-endoplasmic reticulum membrane tethering / autophagosome membrane docking / negative regulation of calcium ion export across plasma membrane / regulation of cardiac muscle cell action potential / presynaptic endocytosis / nitric-oxide synthase binding / regulation of synaptic vesicle exocytosis / filamentous actin / calcineurin-mediated signaling / adenylate cyclase binding / regulation of ryanodine-sensitive calcium-release channel activity / protein phosphatase activator activity / catalytic complex / detection of calcium ion / regulation of synaptic vesicle endocytosis / regulation of cardiac muscle contraction / postsynaptic cytosol / activation of adenylate cyclase activity / cellular response to interferon-beta / phosphatidylinositol 3-kinase binding / calcium channel inhibitor activity / presynaptic cytosol / positive regulation of nitric-oxide synthase activity / vesicle-mediated transport / regulation of release of sequestered calcium ion into cytosol by sarcoplasmic reticulum / enzyme regulator activity / titin binding / sperm midpiece / regulation of calcium-mediated signaling / voltage-gated potassium channel complex / potassium ion transmembrane transport / calcium channel complex / regulation of heart rate / calyx of Held / actin filament organization / response to amphetamine / adenylate cyclase activator activity / sarcomere / protein serine/threonine kinase activator activity / nitric-oxide synthase regulator activity / regulation of cytokinesis / protein localization to plasma membrane / spindle microtubule / positive regulation of receptor signaling pathway via JAK-STAT / calcium-mediated signaling / response to calcium ion / mitochondrial membrane / cellular response to type II interferon 類似検索 - 分子機能 | ||||||
| 生物種 | ![]() ![]() | ||||||
| 手法 | 電子線結晶学 / 電子線トモグラフィー法 / 解像度: 24 Å | ||||||
データ登録者 | Liu, J. / Taylor, D.W. / Krementsova, E.B. / Trybus, K.M. / Taylor, K.A. | ||||||
引用 | ジャーナル: Nature / 年: 2006タイトル: Three-dimensional structure of the myosin V inhibited state by cryoelectron tomography. 著者: Jun Liu / Dianne W Taylor / Elena B Krementsova / Kathleen M Trybus / Kenneth A Taylor / ![]() 要旨: Unconventional myosin V (myoV) is an actin-based molecular motor that has a key function in organelle and mRNA transport, as well as in membrane trafficking. MyoV was the first member of the myosin ...Unconventional myosin V (myoV) is an actin-based molecular motor that has a key function in organelle and mRNA transport, as well as in membrane trafficking. MyoV was the first member of the myosin superfamily shown to be processive, meaning that a single motor protein can 'walk' hand-over-hand along an actin filament for many steps before detaching. Full-length myoV has a low actin-activated MgATPase activity at low [Ca2+], whereas expressed constructs lacking the cargo-binding domain have a high activity regardless of [Ca2+] (refs 5-7). Hydrodynamic data and electron micrographs indicate that the active state is extended, whereas the inactive state is compact. Here we show the first three-dimensional structure of the myoV inactive state. Each myoV molecule consists of two heads that contain an amino-terminal motor domain followed by a lever arm that binds six calmodulins. The heads are followed by a coiled-coil dimerization domain (S2) and a carboxy-terminal globular cargo-binding domain. In the inactive structure, bending of myoV at the head-S2 junction places the cargo-binding domain near the motor domain's ATP-binding pocket, indicating that ATPase inhibition might occur through decreased rates of nucleotide exchange. The actin-binding interfaces are unobstructed, and the lever arm is oriented in a position typical of strong actin-binding states. This structure indicates that motor recycling after cargo delivery might occur through transport on actively treadmilling actin filaments rather than by diffusion. | ||||||
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構造の表示
| ムービー |
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| 構造ビューア | 分子: Molmil Jmol/JSmol |
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ダウンロードとリンク
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ダウンロード
| PDBx/mmCIF形式 | 2dfs.cif.gz | 642.3 KB | 表示 | PDBx/mmCIF形式 |
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| PDB形式 | pdb2dfs.ent.gz | 504.9 KB | 表示 | PDB形式 |
| PDBx/mmJSON形式 | 2dfs.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
| その他 | その他のダウンロード |
-検証レポート
| 文書・要旨 | 2dfs_validation.pdf.gz | 635.8 KB | 表示 | wwPDB検証レポート |
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| 文書・詳細版 | 2dfs_full_validation.pdf.gz | 748.4 KB | 表示 | |
| XML形式データ | 2dfs_validation.xml.gz | 96 KB | 表示 | |
| CIF形式データ | 2dfs_validation.cif.gz | 149.8 KB | 表示 | |
| アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/df/2dfs ftp://data.pdbj.org/pub/pdb/validation_reports/df/2dfs | HTTPS FTP |
-関連構造データ
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リンク
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集合体
| 登録構造単位 | ![]()
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| 単位格子 |
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要素
| #1: タンパク質 | 分子量: 126415.695 Da / 分子数: 2 / 断片: residues 1-1080 / 由来タイプ: 組換発現 / 由来: (組換発現) ![]() 発現宿主: ![]() 参照: UniProt: Q02440 #2: タンパク質 | 分子量: 16721.350 Da / 分子数: 12 / 由来タイプ: 組換発現 / 由来: (組換発現) ![]() 発現宿主: ![]() 参照: UniProt: P62204, UniProt: P0DP26*PLUS |
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-実験情報
-実験
| 実験 | 手法: 電子線結晶学 |
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| EM実験 | 試料の集合状態: TISSUE / 3次元再構成法: 電子線トモグラフィー法 |
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試料調製
| 構成要素 | 名称: Myosin-V / タイプ: TISSUE / 詳細: myosin-V inhibited state |
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| 緩衝液 | 名称: 20mM Na2HPO4, 80-100mM NaCl, 2mM MgCl2, 1mM ADP, 1mM EGTA, 8-9% PEG 8000 詳細: 20mM Na2HPO4, 80-100mM NaCl, 2mM MgCl2, 1mM ADP, 1mM EGTA, 8-9% PEG 8000 |
| 試料 | 濃度: 0.2 mg/ml / 包埋: YES / シャドウイング: NO / 染色: NO / 凍結: NO |
| 試料支持 | 詳細: 200 mesh copper grids covered with a reticulated carbon film |
| 急速凍結 | 装置: HOMEMADE PLUNGER / 凍結剤: ETHANE / 詳細: liquid ethane |
-データ収集
| 顕微鏡 | モデル: FEI/PHILIPS CM300FEG/ST / 日付: 2004年2月1日 |
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| 電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM |
| 電子レンズ | モード: BRIGHT FIELD / 倍率(公称値): 24000 X / 倍率(補正後): 24000 X / 最大 デフォーカス(公称値): 4000 nm / 最小 デフォーカス(公称値): 12000 nm / Cs: 2 mm |
| 試料ホルダ | 温度: 103 K / 傾斜角・最大: 70 ° / 傾斜角・最小: -70 ° |
| 撮影 | 電子線照射量: 30 e/Å2 フィルム・検出器のモデル: TVIPS TEMCAM-F224 (2k x 2k) 詳細: 2048x2048 |
| 放射 | プロトコル: SINGLE WAVELENGTH / 単色(M)・ラウエ(L): M / 散乱光タイプ: electron |
| 放射波長 | 相対比: 1 |
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解析
| EMソフトウェア |
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| CTF補正 | 詳細: CTF gradient correction | |||||||||||||||||||||
| 3次元再構成 | 手法: electron tomography and 3-D average / 解像度: 24 Å / 粒子像の数: 4029 / ピクセルサイズ(公称値): 5.56 Å / ピクセルサイズ(実測値): 5.56 Å 詳細: Tilt series images were aligned using marker-free alignment. Molecular volumes extracted from the cryotomograms, aligned in 3-D. Marker-free alignment is described in the reference: Winkler,H. ...詳細: Tilt series images were aligned using marker-free alignment. Molecular volumes extracted from the cryotomograms, aligned in 3-D. Marker-free alignment is described in the reference: Winkler,H. and Taylor,K.A. Accurate marker-free alignment with simultaneous geometry determination and reconstruction of tilt series in electron tomography. Ultramicroscopy, 106, 240-254 (2006) 対称性のタイプ: 2D CRYSTAL | |||||||||||||||||||||
| 原子モデル構築 | プロトコル: FLEXIBLE FIT / 空間: REAL / Target criteria: Cross correlation 詳細: METHOD--Rigid body docking first and then refined with normal mode flexible fitting. REFINEMENT PROTOCOL--Rigid body docking and normal mode flexible fitting. For the fitting the density ...詳細: METHOD--Rigid body docking first and then refined with normal mode flexible fitting. REFINEMENT PROTOCOL--Rigid body docking and normal mode flexible fitting. For the fitting the density corresponding to the two myosin V heads within the asymmetric unit was segmented from the rest of the map using the watershed transform (Volkman 2002. J.STRUCT.BIOL. 138, 123-129). After fitting the atomic model against the head density, the coordinates were not refined further against the nearest neighbor myosin V heads to remove bad crystal contacts. The alpha-helical coiled-coil domain was manually fit to the and not refined further. | |||||||||||||||||||||
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| 精密化ステップ | サイクル: LAST
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