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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: 9egk | |||||||||
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| タイトル | E3 ubiquitin ligase HUWE1 homolog Tom1p in closed-conformation with internal Acidic Domain deletion | |||||||||
要素 | E3 ubiquitin-protein ligase TOM1 | |||||||||
キーワード | GENE REGULATION / alpha solenoid / E3 ubiquitin ligase / stress response / cell-cycle | |||||||||
| 機能・相同性 | 機能・相同性情報endonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / HECT-type E3 ubiquitin transferase / nucleocytoplasmic transport / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of cell size / nucleus organization / Antigen processing: Ubiquitination & Proteasome degradation / mRNA transport / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) ...endonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / HECT-type E3 ubiquitin transferase / nucleocytoplasmic transport / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of cell size / nucleus organization / Antigen processing: Ubiquitination & Proteasome degradation / mRNA transport / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / Neutrophil degranulation / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / mitotic cell cycle / ubiquitin-dependent protein catabolic process / protein ubiquitination / nucleolus / nucleus / cytoplasm 類似検索 - 分子機能 | |||||||||
| 生物種 | ![]() | |||||||||
| 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: 3.07 Å | |||||||||
データ登録者 | Madrigal, J.M. | |||||||||
| 資金援助 | 米国, 2件
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引用 | ジャーナル: Elife / 年: 2024タイトル: Tom1p ubiquitin ligase structure, interaction with Spt6p, and function in maintaining normal transcript levels and the stability of chromatin in promoters 著者: Madrigal, J. / Schubert, H.L. / Sdano, M.A. / McCullough, L. / Connell, Z. / Formosa, T. / Hill, C.P. | |||||||||
| 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 | 9egk.cif.gz | 933.7 KB | 表示 | PDBx/mmCIF形式 |
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| PDB形式 | pdb9egk.ent.gz | 763.3 KB | 表示 | PDB形式 |
| PDBx/mmJSON形式 | 9egk.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 | 9egk_validation.pdf.gz | 1.2 MB | 表示 | wwPDB検証レポート |
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| 文書・詳細版 | 9egk_full_validation.pdf.gz | 1.2 MB | 表示 | |
| XML形式データ | 9egk_validation.xml.gz | 81 KB | 表示 | |
| CIF形式データ | 9egk_validation.cif.gz | 123 KB | 表示 | |
| アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/eg/9egk ftp://data.pdbj.org/pub/pdb/validation_reports/eg/9egk | 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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要素
| #1: タンパク質 | 分子量: 375512.594 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: E1873-E2131 deleted in construct Precission Protease tag appended to C-terminus 由来: (組換発現) ![]() 遺伝子: TOM1, SSR2, YDR457W, D8035.1 / 発現宿主: ![]() |
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| Has protein modification | 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: 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: Yeast E3 ubiquitin ligase Tom1p with an internal truncation at the acidic domain (residues 1873-2131). タイプ: CELL 詳細: C-terminally tagged with Protein A for purification, cleaved with Prescission Protease. Entity ID: all / 由来: RECOMBINANT |
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| 由来(天然) | 生物種: ![]() |
| 由来(組換発現) | 生物種: ![]() |
| 緩衝液 | pH: 7 |
| 試料 | 濃度: 1 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
| 急速凍結 | 装置: FEI VITROBOT MARK I / 凍結剤: ETHANE / 湿度: 100 % 詳細: Specimens were prepared on Quantifoil R 2/2 Cu300 grids after glow discharge for 1 minute at 25 mA. The detergent sample was concentrated to 8.1 mg/mL in 0.01% CHAPS and 0.05% NP-40. 2.5 uL ...詳細: Specimens were prepared on Quantifoil R 2/2 Cu300 grids after glow discharge for 1 minute at 25 mA. The detergent sample was concentrated to 8.1 mg/mL in 0.01% CHAPS and 0.05% NP-40. 2.5 uL of sample was applied to grids and blotted for 2.5 seconds before vitrification by plunging into liquid ethane. For samples without detergent, protein was concentrated to 0.44 mg/mL and blotted for 1.5 seconds before vitrification. |
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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: OTHER / 最大 デフォーカス(公称値): 2000 nm / 最小 デフォーカス(公称値): 800 nm |
| 撮影 | 電子線照射量: 47 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 実像数: 7236 詳細: A total of 2,865 movies were recorded from grids without detergent at 81,000x magnification on a 300 kV Titan Krios G3 electron microscope with a K3 direct detector (nominal resolution after ...詳細: A total of 2,865 movies were recorded from grids without detergent at 81,000x magnification on a 300 kV Titan Krios G3 electron microscope with a K3 direct detector (nominal resolution after 2x binning is 1.06A/px). Electron exposure was 47 (e-/A2) over a total of 40 frames. A total of 4,371 movies were recorded from with-detergent grids using the same electron microscope and collection parameters. |
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解析
| EMソフトウェア |
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| CTF補正 | タイプ: NONE | ||||||||||||||||||||||||
| 3次元再構成 | 解像度: 3.07 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 462711 詳細: The separately processed dataset #1 and #2 particles were combined for a total of 531,549 starting particles for further processing and refinement. A round of 2D classification resulted in ...詳細: The separately processed dataset #1 and #2 particles were combined for a total of 531,549 starting particles for further processing and refinement. A round of 2D classification resulted in selection of 477,889 particles, which were Fourier cropped from a box size of 680 to 320 pixels for further processing. Two-component 3DVA was performed after map refinement of the combined dataset with a filter resolution of 4 A. 3DVA display was subsequently run to give 8 intermediates along the first principal component at a filter resolution of 5 A. The top 4 intermediates with the highest number of particles were combined to yield 462,711 and refined using non-uniform refinement to an estimated overall resolution of 3.07 A. 対称性のタイプ: POINT | ||||||||||||||||||||||||
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万見について






米国, 2件
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