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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: 9i1j | ||||||
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| タイトル | Cryo-EM structure of mouse RNF213:UBE2L3 transthiolation intermediate, chemically stabilized, and ATPgS | ||||||
要素 |
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キーワード | LIGASE / E3 ligase / ubiquitin / AAA ATPase | ||||||
| 機能・相同性 | 機能・相同性情報lipid ubiquitination / negative regulation of non-canonical Wnt signaling pathway / lipid droplet formation / cell cycle phase transition / ubiquitin-protein transferase activator activity / xenophagy / protein K11-linked ubiquitination / Antigen processing: Ubiquitination & Proteasome degradation / sprouting angiogenesis / 転移酵素; アシル基を移すもの; アミノアシル基を移すもの ...lipid ubiquitination / negative regulation of non-canonical Wnt signaling pathway / lipid droplet formation / cell cycle phase transition / ubiquitin-protein transferase activator activity / xenophagy / protein K11-linked ubiquitination / Antigen processing: Ubiquitination & Proteasome degradation / sprouting angiogenesis / 転移酵素; アシル基を移すもの; アミノアシル基を移すもの / : / cellular response to glucocorticoid stimulus / E2 ubiquitin-conjugating enzyme / cellular response to steroid hormone stimulus / ubiquitin conjugating enzyme activity / regulation of lipid metabolic process / protein K63-linked ubiquitination / immune system process / ubiquitin ligase complex / protein autoubiquitination / lipid droplet / Synthesis of active ubiquitin: roles of E1 and E2 enzymes / positive regulation of protein ubiquitination / PINK1-PRKN Mediated Mitophagy / Regulation of TNFR1 signaling / protein modification process / RING-type E3 ubiquitin transferase / Regulation of necroptotic cell death / 加水分解酵素; 酸無水物に作用; 酸無水物に作用・細胞または細胞小器官の運動に関与 / protein polyubiquitination / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / Antigen processing: Ubiquitination & Proteasome degradation / E3 ubiquitin ligases ubiquitinate target proteins / angiogenesis / ubiquitin-dependent protein catabolic process / transcription coactivator activity / cell population proliferation / defense response to bacterium / protein ubiquitination / ubiquitin protein ligase binding / regulation of DNA-templated transcription / enzyme binding / ATP hydrolysis activity / RNA binding / zinc ion binding / nucleoplasm / ATP binding / nucleus / cytosol / cytoplasm 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: 3.8 Å | ||||||
データ登録者 | Grabarczyk, D.B. / Ahel, J. / Clausen, T. | ||||||
| 資金援助 | オーストリア, 1件
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引用 | ジャーナル: Nat Commun / 年: 2025タイトル: ATP functions as a pathogen-associated molecular pattern to activate the E3 ubiquitin ligase RNF213. 著者: Juraj Ahel / Arda Balci / Victoria Faas / Daniel B Grabarczyk / Roosa Harmo / Daniel R Squair / Jiazhen Zhang / Elisabeth Roitinger / Frederic Lamoliatte / Sunil Mathur / Luiza Deszcz / ...著者: Juraj Ahel / Arda Balci / Victoria Faas / Daniel B Grabarczyk / Roosa Harmo / Daniel R Squair / Jiazhen Zhang / Elisabeth Roitinger / Frederic Lamoliatte / Sunil Mathur / Luiza Deszcz / Lillie E Bell / Anita Lehner / Thomas L Williams / Hanna Sowar / Anton Meinhart / Nicola T Wood / Tim Clausen / Satpal Virdee / Adam J Fletcher / ![]() 要旨: The giant E3 ubiquitin ligase RNF213 is a conserved component of mammalian cell-autonomous immunity, limiting the replication of bacteria, viruses and parasites. To understand how RNF213 reacts to ...The giant E3 ubiquitin ligase RNF213 is a conserved component of mammalian cell-autonomous immunity, limiting the replication of bacteria, viruses and parasites. To understand how RNF213 reacts to these unrelated pathogens, we employ chemical and structural biology to find that ATP binding to its ATPases Associated with diverse cellular Activities (AAA) core activates its E3 function. We develop methodology for proteome-wide E3 activity profiling inside living cells, revealing that RNF213 undergoes a reversible switch in E3 activity in response to cellular ATP abundance. Interferon stimulation of macrophages raises intracellular ATP levels and primes RNF213 E3 activity, while glycolysis inhibition depletes ATP and downregulates E3 activity. These data imply that ATP bears hallmarks of a danger/pathogen associated molecular pattern, coordinating cell-autonomous defence. Furthermore, quantitative labelling of RNF213 with E3-activity probes enabled us to identify the catalytic cysteine required for substrate ubiquitination and obtain a cryo-EM structure of the RNF213-E2-ubiquitin conjugation enzyme transfer intermediate, illuminating an unannotated E2 docking site. Together, our data demonstrate that RNF213 represents a new class of ATP-dependent E3 enzyme, employing distinct catalytic and regulatory mechanisms adapted to its specialised role in the broad defence against intracellular pathogens. #1: ジャーナル: Acta Crystallogr D Struct Biol / 年: 2019 タイトル: Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. 著者: Dorothee Liebschner / Pavel V Afonine / Matthew L Baker / Gábor Bunkóczi / Vincent B Chen / Tristan I Croll / Bradley Hintze / Li Wei Hung / Swati Jain / Airlie J McCoy / Nigel W Moriarty / ...著者: Dorothee Liebschner / Pavel V Afonine / Matthew L Baker / Gábor Bunkóczi / Vincent B Chen / Tristan I Croll / Bradley Hintze / Li Wei Hung / Swati Jain / Airlie J McCoy / Nigel W Moriarty / Robert D Oeffner / Billy K Poon / Michael G Prisant / Randy J Read / Jane S Richardson / David C Richardson / Massimo D Sammito / Oleg V Sobolev / Duncan H Stockwell / Thomas C Terwilliger / Alexandre G Urzhumtsev / Lizbeth L Videau / Christopher J Williams / Paul D Adams / ![]() 要旨: Diffraction (X-ray, neutron and electron) and electron cryo-microscopy are powerful methods to determine three-dimensional macromolecular structures, which are required to understand biological ...Diffraction (X-ray, neutron and electron) and electron cryo-microscopy are powerful methods to determine three-dimensional macromolecular structures, which are required to understand biological processes and to develop new therapeutics against diseases. The overall structure-solution workflow is similar for these techniques, but nuances exist because the properties of the reduced experimental data are different. Software tools for structure determination should therefore be tailored for each method. Phenix is a comprehensive software package for macromolecular structure determination that handles data from any of these techniques. Tasks performed with Phenix include data-quality assessment, map improvement, model building, the validation/rebuilding/refinement cycle and deposition. Each tool caters to the type of experimental data. The design of Phenix emphasizes the automation of procedures, where possible, to minimize repetitive and time-consuming manual tasks, while default parameters are chosen to encourage best practice. A graphical user interface provides access to many command-line features of Phenix and streamlines the transition between programs, project tracking and re-running of previous tasks. | ||||||
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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 | 9i1j.cif.gz | 838.8 KB | 表示 | PDBx/mmCIF形式 |
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| PDB形式 | pdb9i1j.ent.gz | 665.5 KB | 表示 | PDB形式 |
| PDBx/mmJSON形式 | 9i1j.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 | 9i1j_validation.pdf.gz | 1.8 MB | 表示 | wwPDB検証レポート |
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| 文書・詳細版 | 9i1j_full_validation.pdf.gz | 1.9 MB | 表示 | |
| XML形式データ | 9i1j_validation.xml.gz | 135.9 KB | 表示 | |
| CIF形式データ | 9i1j_validation.cif.gz | 205.7 KB | 表示 | |
| アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/i1/9i1j ftp://data.pdbj.org/pub/pdb/validation_reports/i1/9i1j | HTTPS FTP |
-関連構造データ
| 関連構造データ | ![]() 52571MC ![]() 9i1iC 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
-タンパク質 , 2種, 2分子 AB
| #1: タンパク質 | 分子量: 552623.375 Da / 分子数: 1 / 変異: delta N300 / 由来タイプ: 組換発現 / 由来: (組換発現) ![]() Trichoplusia ni (イラクサキンウワバ)参照: UniProt: E9Q555, RING-type E3 ubiquitin transferase, 加水分解酵素; 酸無水物に作用; 酸無水物に作用・細胞または細胞小器官の運動に関与 |
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| #2: タンパク質 | 分子量: 19197.908 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: UBE2L3, UBCE7, UBCH7 / 発現宿主: ![]() |
-非ポリマー , 4種, 6分子 






| #3: 化合物 | ChemComp-ATP / | ||
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| #4: 化合物 | ChemComp-MG / | ||
| #5: 化合物 | | #6: 化合物 | |
-詳細
| 研究の焦点であるリガンドがあるか | Y |
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| Has protein modification | Y |
-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: Mouse RNF213:UBE2L3 transthiolation intermediate / タイプ: COMPLEX / Entity ID: #1-#2 / 由来: RECOMBINANT |
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| 由来(天然) | 生物種: ![]() |
| 由来(組換発現) | 生物種: Trichoplusia ni (イラクサキンウワバ) |
| 緩衝液 | pH: 8 |
| 試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
| 急速凍結 | 凍結剤: ETHANE |
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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: 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 / 最大 デフォーカス(公称値): 2500 nm / 最小 デフォーカス(公称値): 800 nm |
| 撮影 | 電子線照射量: 50 e/Å2 フィルム・検出器のモデル: FEI FALCON IV (4k x 4k) |
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解析
| EMソフトウェア | 名称: PHENIX / バージョン: 1.21.2_5419 / カテゴリ: モデル精密化 |
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| CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION |
| 3次元再構成 | 解像度: 3.8 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 31607 / 対称性のタイプ: POINT |
| 精密化 | 最高解像度: 3.8 Å 立体化学のターゲット値: REAL-SPACE (WEIGHTED MAP SUM AT ATOM CENTERS) |
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万見について





Homo sapiens (ヒト)
オーストリア, 1件
引用




PDBj





Trichoplusia ni (イラクサキンウワバ)

FIELD EMISSION GUN