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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: 8a3d | ||||||
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| タイトル | Human mature large subunit of the ribosome with eIF6 and homoharringtonine bound | ||||||
要素 |
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キーワード | TRANSLATION / Ribosome / human / rna modifications / water / magnesium / potassium / homoharringtonine | ||||||
| 機能・相同性 | 機能・相同性情報lamin filament / regulation of fatty acid biosynthetic process / regulation of megakaryocyte differentiation / miRNA-mediated post-transcriptional gene silencing / miRNA-mediated gene silencing by inhibition of translation / translation at presynapse / embryonic brain development / exit from mitosis / eukaryotic 80S initiation complex / negative regulation of protein neddylation ...lamin filament / regulation of fatty acid biosynthetic process / regulation of megakaryocyte differentiation / miRNA-mediated post-transcriptional gene silencing / miRNA-mediated gene silencing by inhibition of translation / translation at presynapse / embryonic brain development / exit from mitosis / eukaryotic 80S initiation complex / negative regulation of protein neddylation / optic nerve development / regulation of translation involved in cellular response to UV / axial mesoderm development / negative regulation of formation of translation preinitiation complex / regulation of G1 to G0 transition / ribosomal protein import into nucleus / protein-DNA complex disassembly / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / 90S preribosome assembly / retinal ganglion cell axon guidance / GAIT complex / positive regulation of DNA damage response, signal transduction by p53 class mediator / TORC2 complex binding / alpha-beta T cell differentiation / G1 to G0 transition / regulation of glycolytic process / middle ear morphogenesis / regulation of reactive oxygen species metabolic process / cytoplasmic side of rough endoplasmic reticulum membrane / maturation of 5.8S rRNA / negative regulation of ubiquitin protein ligase activity / homeostatic process / macrophage chemotaxis / lung morphogenesis / ribosomal large subunit binding / positive regulation of natural killer cell proliferation / Protein hydroxylation / Peptide chain elongation / Selenocysteine synthesis / Formation of a pool of free 40S subunits / Eukaryotic Translation Termination / ubiquitin ligase inhibitor activity / cellular response to actinomycin D / Response of EIF2AK4 (GCN2) to amino acid deficiency / blastocyst development / SRP-dependent cotranslational protein targeting to membrane / negative regulation of ubiquitin-dependent protein catabolic process / positive regulation of signal transduction by p53 class mediator / Viral mRNA Translation / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / GTP hydrolysis and joining of the 60S ribosomal subunit / protein localization to nucleus / L13a-mediated translational silencing of Ceruloplasmin expression / positive regulation of protein binding / Major pathway of rRNA processing in the nucleolus and cytosol / protein targeting / protein-RNA complex assembly / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / ribosomal subunit export from nucleus / maturation of LSU-rRNA / rough endoplasmic reticulum / translation regulator activity / MDM2/MDM4 family protein binding / translation initiation factor activity / negative regulation of proteasomal ubiquitin-dependent protein catabolic process / embryo implantation / cytosolic ribosome / cellular response to interleukin-4 / ossification / assembly of large subunit precursor of preribosome / positive regulation of translation / regulation of signal transduction by p53 class mediator / ribosomal large subunit biogenesis / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / mRNA 3'-UTR binding / cytosolic ribosome assembly / innate immune response in mucosa / skeletal system development / DNA damage response, signal transduction by p53 class mediator / sensory perception of sound / cellular response to gamma radiation / response to insulin / bone development / mRNA 5'-UTR binding / cellular response to type II interferon / Regulation of expression of SLITs and ROBOs / transcription coactivator binding / cytoplasmic ribonucleoprotein granule / rRNA processing / antimicrobial humoral immune response mediated by antimicrobial peptide / cellular response to UV / antibacterial humoral response / presynapse / heparin binding / large ribosomal subunit / regulation of translation / ribosome binding / retina development in camera-type eye / cell body / 5S rRNA binding 類似検索 - 分子機能 | ||||||
| 生物種 | Homo sapiens (ヒト) | ||||||
| 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: 1.67 Å | ||||||
データ登録者 | Faille, A. / Warren, A.J. / Dent, K.C. | ||||||
| 資金援助 | 英国, 1件
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引用 | ジャーナル: bioRxiv / 年: 2023タイトル: The chemical landscape of the human ribosome at 1.67 Å resolution. 著者: Alexandre Faille / Kyle C Dent / Simone Pellegrino / Pekka Jaako / Alan J Warren 要旨: The ability of ribosomes to translate the genetic code into protein requires a finely tuned ion and solvent ecosystem. However, the lack of high-resolution structures has precluded accurate ...The ability of ribosomes to translate the genetic code into protein requires a finely tuned ion and solvent ecosystem. However, the lack of high-resolution structures has precluded accurate positioning of all the functional elements of the ribosome and limited our understanding of the specific role of ribosomal RNA chemical modifications in modulating ribosome function in health and disease. Here, using a new sample preparation methodology based on functionalised pristine graphene-coated grids, we solve the cryo-EM structure of the human large ribosomal subunit to a resolution of 1.67 Å. The accurate assignment of water molecules, magnesium and potassium ions in our model highlights the fundamental biological role of ribosomal RNA methylation in harnessing unconventional carbon-oxygen hydrogen bonds to establish chemical interactions with the environment and fine-tune the functional interplay with tRNA. In addition, the structures of three translational inhibitors bound to the human large ribosomal subunit at better than 2 Å resolution provide mechanistic insights into how three key druggable pockets of the ribosome are targeted and illustrate the potential of this methodology to accelerate high-throughput structure-based design of anti-cancer therapeutics. | ||||||
| 履歴 |
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構造の表示
| 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 | 8a3d.cif.gz | 3.7 MB | 表示 | PDBx/mmCIF形式 |
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| PDB形式 | pdb8a3d.ent.gz | 表示 | PDB形式 | |
| PDBx/mmJSON形式 | 8a3d.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 | 8a3d_validation.pdf.gz | 1.6 MB | 表示 | wwPDB検証レポート |
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| 文書・詳細版 | 8a3d_full_validation.pdf.gz | 1.8 MB | 表示 | |
| XML形式データ | 8a3d_validation.xml.gz | 239.8 KB | 表示 | |
| CIF形式データ | 8a3d_validation.cif.gz | 459.9 KB | 表示 | |
| アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/a3/8a3d ftp://data.pdbj.org/pub/pdb/validation_reports/a3/8a3d | HTTPS FTP |
-関連構造データ
| 関連構造データ | ![]() 15113MC 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 F&H 検索 |
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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 , 3種, 3分子 ABC
| #1: RNA鎖 | 分子量: 1640812.500 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 / 参照: GenBank: 86475748 |
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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: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 / 参照: GenBank: 23898 |
| #3: RNA鎖 | 分子量: 50463.840 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 / 参照: GenBank: 555853 |
+60S ribosomal protein ... , 39種, 39分子 DEFGHmnopqrslIJKLMNOPQRSTUVWXY...
-タンパク質 , 3種, 3分子 bgh
| #36: タンパク質 | 分子量: 21730.748 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 |
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| #41: タンパク質 | 分子量: 11470.713 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 / 参照: UniProt: A0A3Q0DRJ6 |
| #45: タンパク質 | 分子量: 26620.010 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 / 参照: UniProt: P56537 |
-非ポリマー , 8種, 12367分子 














| #46: 化合物 | ChemComp-MG / #47: 化合物 | ChemComp-K / #48: 化合物 | #49: 化合物 | ChemComp-HMT / ( | #50: 化合物 | ChemComp-SPM / | #51: 化合物 | ChemComp-EPE / | #52: 化合物 | ChemComp-ZN / #53: 水 | ChemComp-HOH / | |
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-詳細
| 研究の焦点であるリガンドがあるか | N |
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-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: Human mature large subunit of the ribosome with eIF6 bound タイプ: RIBOSOME / Entity ID: #1-#45 / 由来: NATURAL | |||||||||||||||||||||||||
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| 分子量 | 実験値: NO | |||||||||||||||||||||||||
| 由来(天然) | 生物種: Homo sapiens (ヒト) | |||||||||||||||||||||||||
| 緩衝液 | pH: 7.5 | |||||||||||||||||||||||||
| 緩衝液成分 |
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| 試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | |||||||||||||||||||||||||
| 試料支持 | 詳細: Grid was functionalised with 1-pyrenemethylamine. See methods section in published article. グリッドの材料: COPPER / グリッドのサイズ: 300 divisions/in. / グリッドのタイプ: Quantifoil R2/2 | |||||||||||||||||||||||||
| 急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 277 K / 詳細: 30 seconds 'wait' time |
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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 |
| 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): 105000 X / 最大 デフォーカス(公称値): 1800 nm / 最小 デフォーカス(公称値): 1000 nm / Cs: 2.7 mm / C2レンズ絞り径: 50 µm / アライメント法: COMA FREE |
| 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: 2.2 sec. / 電子線照射量: 51 e/Å2 フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 撮影したグリッド数: 1 / 実像数: 9973 |
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解析
| ソフトウェア | 名称: REFMAC / バージョン: 5.8.0267 / 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 AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||
| 粒子像の選択 | 選択した粒子像数: 1147785 | ||||||||||||||||||||||||||||||||||||
| 3次元再構成 | 解像度: 1.67 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 1088709 / クラス平均像の数: 1 / 対称性のタイプ: POINT | ||||||||||||||||||||||||||||||||||||
| 原子モデル構築 | B value: 40 / プロトコル: AB INITIO MODEL / 空間: REAL | ||||||||||||||||||||||||||||||||||||
| 原子モデル構築 | PDB-ID: 6EK0![]() 6ek0 Accession code: 6EK0 / Source name: PDB / タイプ: experimental model | ||||||||||||||||||||||||||||||||||||
| 精密化 | 解像度: 1.67→270.25 Å / Cor.coef. Fo:Fc: 0.688
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| 原子変位パラメータ | Biso mean: 35.442 Å2 | ||||||||||||||||||||||||||||||||||||
| LS精密化 シェル | 最高解像度: 1.67 Å /
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万見について




Homo sapiens (ヒト)
英国, 1件
引用
PDBj






































FIELD EMISSION GUN
