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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: 5aj0 | ||||||
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| タイトル | Cryo electron microscopy of actively translating human polysomes (POST state). | ||||||
要素 |
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キーワード | RIBOSOME / MAMMALIAN RIBOSOME / TRANSLATION / POLYSOME / CRYO ELECTRON MICROSCOPY / ELONGATION CYCLE | ||||||
| 機能・相同性 | 機能・相同性情報embryonic brain development / translation at presynapse / exit from mitosis / optic nerve development / response to insecticide / regulation of translation involved in cellular response to UV / eukaryotic 80S initiation complex / negative regulation of formation of translation preinitiation complex / axial mesoderm development / negative regulation of endoplasmic reticulum unfolded protein response ...embryonic brain development / translation at presynapse / exit from mitosis / optic nerve development / response to insecticide / regulation of translation involved in cellular response to UV / eukaryotic 80S initiation complex / negative regulation of formation of translation preinitiation complex / axial mesoderm development / negative regulation of endoplasmic reticulum unfolded protein response / ribosomal protein import into nucleus / regulation of G1 to G0 transition / retinal ganglion cell axon guidance / oxidized pyrimidine DNA binding / response to TNF agonist / positive regulation of base-excision repair / protein-DNA complex disassembly / positive regulation of ubiquitin-protein transferase activity / positive regulation of respiratory burst involved in inflammatory response / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / positive regulation of gastrulation / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage / protein tyrosine kinase inhibitor activity / 90S preribosome assembly / IRE1-RACK1-PP2A complex / positive regulation of Golgi to plasma membrane protein transport / nucleolus organization / alpha-beta T cell differentiation / positive regulation of DNA-templated transcription initiation / TNFR1-mediated ceramide production / positive regulation of DNA damage response, signal transduction by p53 class mediator / GAIT complex / negative regulation of RNA splicing / TORC2 complex binding / neural crest cell differentiation / supercoiled DNA binding / NF-kappaB complex / negative regulation of DNA repair / G1 to G0 transition / cytoplasmic translational initiation / oxidized purine DNA binding / cysteine-type endopeptidase activator activity involved in apoptotic process / middle ear morphogenesis / negative regulation of intrinsic apoptotic signaling pathway in response to hydrogen peroxide / rRNA modification in the nucleus and cytosol / negative regulation of bicellular tight junction assembly / ubiquitin-like protein conjugating enzyme binding / regulation of establishment of cell polarity / negative regulation of phagocytosis / Formation of the ternary complex, and subsequently, the 43S complex / erythrocyte homeostasis / cytoplasmic side of rough endoplasmic reticulum membrane / protein kinase A binding / Ribosomal scanning and start codon recognition / ion channel inhibitor activity / laminin receptor activity / Translation initiation complex formation / homeostatic process / pigmentation / positive regulation of mitochondrial depolarization / macrophage chemotaxis / lung morphogenesis / negative regulation of Wnt signaling pathway / positive regulation of natural killer cell proliferation / fibroblast growth factor binding / monocyte chemotaxis / BH3 domain binding / Protein hydroxylation / negative regulation of translational frameshifting / SARS-CoV-1 modulates host translation machinery / regulation of adenylate cyclase-activating G protein-coupled receptor signaling pathway / TOR signaling / positive regulation of GTPase activity / iron-sulfur cluster binding / mTORC1-mediated signalling / Peptide chain elongation / regulation of cell division / Selenocysteine synthesis / Formation of a pool of free 40S subunits / cellular response to ethanol / blastocyst development / Eukaryotic Translation Termination / positive regulation of intrinsic apoptotic signaling pathway by p53 class mediator / SRP-dependent cotranslational protein targeting to membrane / Response of EIF2AK4 (GCN2) to amino acid deficiency / negative regulation of protein binding / protein serine/threonine kinase inhibitor activity / Viral mRNA Translation / ubiquitin ligase inhibitor activity / negative regulation of respiratory burst involved in inflammatory response / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / positive regulation of signal transduction by p53 class mediator / GTP hydrolysis and joining of the 60S ribosomal subunit / protein localization to nucleus / negative regulation of ubiquitin-dependent protein catabolic process / L13a-mediated translational silencing of Ceruloplasmin expression / Major pathway of rRNA processing in the nucleolus and cytosol / protein targeting / regulation of translational fidelity 類似検索 - 分子機能 | ||||||
| 生物種 | Homo sapiens (ヒト) Yersinia pseudotuberculosis (仮性結核菌) | ||||||
| 手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.5 Å | ||||||
データ登録者 | Behrmann, E. / Loerke, J. / Budkevich, T.V. / Yamamoto, K. / Schmidt, A. / Penczek, P.A. / Vos, M.R. / Burger, J. / Mielke, T. / Scheerer, P. / Spahn, C.M.T. | ||||||
引用 | ジャーナル: Cell / 年: 2015タイトル: Structural snapshots of actively translating human ribosomes. 著者: Elmar Behrmann / Justus Loerke / Tatyana V Budkevich / Kaori Yamamoto / Andrea Schmidt / Pawel A Penczek / Matthijn R Vos / Jörg Bürger / Thorsten Mielke / Patrick Scheerer / Christian M T Spahn / ![]() 要旨: Macromolecular machines, such as the ribosome, undergo large-scale conformational changes during their functional cycles. Although their mode of action is often compared to that of mechanical ...Macromolecular machines, such as the ribosome, undergo large-scale conformational changes during their functional cycles. Although their mode of action is often compared to that of mechanical machines, a crucial difference is that, at the molecular dimension, thermodynamic effects dominate functional cycles, with proteins fluctuating stochastically between functional states defined by energetic minima on an energy landscape. Here, we have used cryo-electron microscopy to image ex-vivo-derived human polysomes as a source of actively translating ribosomes. Multiparticle refinement and 3D variability analysis allowed us to visualize a variety of native translation intermediates. Significantly populated states include not only elongation cycle intermediates in pre- and post-translocational states, but also eEF1A-containing decoding and termination/recycling complexes. Focusing on the post-translocational state, we extended this assessment to the single-residue level, uncovering striking details of ribosome-ligand interactions and identifying both static and functionally important dynamic elements. | ||||||
| 履歴 |
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| Remark 700 | SHEET DETERMINATION METHOD: DSSP THE SHEETS PRESENTED AS "BA" IN EACH CHAIN ON SHEET RECORDS BELOW ...SHEET DETERMINATION METHOD: DSSP THE SHEETS PRESENTED AS "BA" IN EACH CHAIN ON SHEET RECORDS BELOW IS ACTUALLY AN 6-STRANDED BARREL THIS IS REPRESENTED BY A 7-STRANDED SHEET IN WHICH THE FIRST AND LAST STRANDS ARE IDENTICAL. SHEET DETERMINATION METHOD: DSSP THE SHEETS PRESENTED AS "VB" IN EACH CHAIN ON SHEET RECORDS BELOW IS ACTUALLY AN 5-STRANDED BARREL THIS IS REPRESENTED BY A 6-STRANDED SHEET IN WHICH THE FIRST AND LAST STRANDS ARE IDENTICAL. SHEET DETERMINATION METHOD: DSSP THE SHEETS PRESENTED AS "FA" IN EACH CHAIN ON SHEET RECORDS BELOW IS ACTUALLY AN 6-STRANDED BARREL THIS IS REPRESENTED BY A 7-STRANDED SHEET IN WHICH THE FIRST AND LAST STRANDS ARE IDENTICAL. |
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構造の表示
| ムービー |
ムービービューア |
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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 | 5aj0.cif.gz | 4.8 MB | 表示 | PDBx/mmCIF形式 |
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| PDB形式 | pdb5aj0.ent.gz | 表示 | PDB形式 | |
| PDBx/mmJSON形式 | 5aj0.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
| アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/aj/5aj0 ftp://data.pdbj.org/pub/pdb/validation_reports/aj/5aj0 | HTTPS FTP |
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-関連構造データ
| 関連構造データ | ![]() 2875MC ![]() 2902C ![]() 2903C ![]() 2904C ![]() 2905C ![]() 2906C ![]() 2907C ![]() 2908C ![]() 2909C ![]() 2910C ![]() 2911C 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 |
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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
-RNA鎖 , 6種, 7分子 A3A4A2B1BvBwBx
| #1: RNA鎖 | 分子量: 62616.344 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK 293T / 参照: GenBank: 51477016 | ||
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| #2: RNA鎖 | 分子量: 38998.078 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK 293T / 参照: GenBank: 23898 | ||
| #48: RNA鎖 | 分子量: 1627178.625 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK 293T / 参照: GenBank: 337381 | ||
| #49: RNA鎖 | 分子量: 602776.875 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK 293T / 参照: GenBank: 36162 | ||
| #83: RNA鎖 | 分子量: 24485.539 Da / 分子数: 2 / 由来タイプ: 天然 / 詳細: MIXTURE OF ALL ENDOGENOUS PRESENT 由来: (天然) Yersinia pseudotuberculosis (仮性結核菌)細胞株: HEK 293T / 参照: GenBank: 755367513 #84: RNA鎖 | | 分子量: 8527.686 Da / 分子数: 1 / 由来タイプ: 天然 / 詳細: MIXTURE OF ALL ENDOGENOUS PRESENT / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK 293T |
+60S ribosomal protein ... , 43種, 43分子 AAABACADAEAFAGAHAIAJALAMANAOAPAQARASATAUAVAWAXAYAZAaAbAcAdAe...
-タンパク質 , 4種, 4分子 AKAmBfBg
| #13: タンパク質 | 分子量: 34309.418 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK 293T / 参照: UniProt: P05388 |
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| #41: タンパク質 | 分子量: 14758.394 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK 293T / 参照: UniProt: P62987 |
| #81: タンパク質 | 分子量: 18004.041 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK 293T / 参照: UniProt: P62979 |
| #82: タンパク質 | 分子量: 35115.652 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK 293T / 参照: UniProt: P63244 |
+40S ribosomal protein ... , 31種, 31分子 BABBBCBDBEBFBGBHBIBJBKBLBMBNBOBPBQBRBSBTBUBVBWBXBYBZBaBbBcBdBe
-タンパク質・ペプチド , 1種, 1分子 By
| #85: タンパク質・ペプチド | 分子量: 2060.531 Da / 分子数: 1 / 由来タイプ: 天然 / 詳細: MIXTURE OF ALL ENDOGENOUS PRESENT / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK 293T |
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-非ポリマー , 2種, 331分子 


| #86: 化合物 | ChemComp-MG / #87: 化合物 | ChemComp-ZN / |
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-詳細
| Has protein modification | Y |
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| 配列の詳細 | CGCGACCUCA |
-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: PARTICLE / 3次元再構成法: 単粒子再構成法 |
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試料調製
| 構成要素 | 名称: NATIVE RIBOSOMAL COMPLEX FROM POLYSOMES - POST STATE タイプ: RIBOSOME |
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| 緩衝液 | 名称: 20 MM HEPES-KOH, PH 7.5, 100 MM KCL, 1.5 MM MGCL2, 0.5 MM SPERMIDINE, 0.04 MM SPERMINE, 1 MM DTT pH: 7.5 詳細: 20 MM HEPES-KOH, PH 7.5, 100 MM KCL, 1.5 MM MGCL2, 0.5 MM SPERMIDINE, 0.04 MM SPERMINE, 1 MM DTT |
| 試料 | 濃度: 3.5 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
| 試料支持 | 詳細: OTHER |
| 急速凍結 | 装置: FEI VITROBOT MARK II / 凍結剤: ETHANE 詳細: VITRIFICATION 1 -- CRYOGEN- ETHANE, HUMIDITY- 100, TEMPERATURE- 93, INSTRUMENT- FEI VITROBOT MARK II, METHOD- BLOT FOR 2-4 SECONDS BEFORE PLUNGING, |
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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: FEI TITAN KRIOS / 日付: 2012年8月20日 / 詳細: DATA WAS COLLECTED AUTOMATICALLY WITH LEGINON |
| 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): 115000 X / 倍率(補正後): 205000 X / 最大 デフォーカス(公称値): 4500 nm / 最小 デフォーカス(公称値): 2000 nm / Cs: 2 mm |
| 撮影 | 電子線照射量: 20 e/Å2 フィルム・検出器のモデル: TVIPS TEMCAM-F416 (4k x 4k) |
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解析
| EMソフトウェア |
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| CTF補正 | 詳細: DEFOCUS GROUPS | ||||||||||||||||||||
| 対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||
| 3次元再構成 | 手法: MULTI-REFERENCE TEMPLATE MATCHING / 解像度: 3.5 Å / 粒子像の数: 313321 / ピクセルサイズ(実測値): 0.945 Å 倍率補正: CROSS- -CORRELATION DENSITIES WITHIN SPHERICAL SHELL 詳細: SUBMISSION BASED ON EXPERIMENTAL DATA FROM EMDB EMD-2875. (DEPOSITION ID: 13060). 対称性のタイプ: POINT | ||||||||||||||||||||
| 原子モデル構築 | 空間: REAL | ||||||||||||||||||||
| 原子モデル構築 | PDB-ID: 4UJE Accession code: 4UJE / Source name: PDB / タイプ: experimental model | ||||||||||||||||||||
| 精密化 | 最高解像度: 3.5 Å | ||||||||||||||||||||
| 精密化ステップ | サイクル: LAST / 最高解像度: 3.5 Å
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コントローラー
万見について




Homo sapiens (ヒト)
Yersinia pseudotuberculosis (仮性結核菌)
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FIELD EMISSION GUN
