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基本情報
登録情報 | データベース: EMDB / ID: EMD-3136 | |||||||||
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タイトル | A Single Alpha Helix Drives Extensive Remodeling of the Proteasome Lid and Completion of Regulatory Particle Assembly | |||||||||
![]() | Reconstruction of LP2 | |||||||||
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![]() | proteasome / ubiquitin / proteolysis / electron microscopy / mass spectrometry | |||||||||
機能・相同性 | ![]() protein deneddylation => GO:0000338 / molecular_function / SAGA complex localization to transcription regulatory region / Metalloprotease DUBs / peroxisome fission / proteasome storage granule assembly / transcription export complex 2 / protein deneddylation / maintenance of DNA trinucleotide repeats / filamentous growth ...protein deneddylation => GO:0000338 / molecular_function / SAGA complex localization to transcription regulatory region / Metalloprotease DUBs / peroxisome fission / proteasome storage granule assembly / transcription export complex 2 / protein deneddylation / maintenance of DNA trinucleotide repeats / filamentous growth / COP9 signalosome / proteasome regulatory particle / proteasome regulatory particle, lid subcomplex / mitochondrial fission / metal-dependent deubiquitinase activity / Cross-presentation of soluble exogenous antigens (endosomes) / TNFR2 non-canonical NF-kB pathway / Ubiquitin Mediated Degradation of Phosphorylated Cdc25A / Regulation of PTEN stability and activity / KEAP1-NFE2L2 pathway / CDK-mediated phosphorylation and removal of Cdc6 / Neddylation / FBXL7 down-regulates AURKA during mitotic entry and in early mitosis / proteasome binding / regulation of protein catabolic process / Orc1 removal from chromatin / protein deubiquitination / MAPK6/MAPK4 signaling / proteasome storage granule / Antigen processing: Ubiquitination & Proteasome degradation / proteasome assembly / Ub-specific processing proteases / mRNA export from nucleus / enzyme regulator activity / protein folding chaperone / Neutrophil degranulation / proteasome complex / double-strand break repair via homologous recombination / metallopeptidase activity / ubiquitin-dependent protein catabolic process / proteasome-mediated ubiquitin-dependent protein catabolic process / ubiquitinyl hydrolase 1 / cysteine-type deubiquitinase activity / molecular adaptor activity / regulation of cell cycle / structural molecule activity / positive regulation of transcription by RNA polymerase II / mitochondrion / nucleus / metal ion binding / cytoplasm / cytosol 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / ネガティブ染色法 / 解像度: 16.0 Å | |||||||||
![]() | Tomko Jr RJ / Taylor DW / Chen ZA / Wang HW / Rappsilber J / Hochstrasser M | |||||||||
![]() | ![]() タイトル: A Single α Helix Drives Extensive Remodeling of the Proteasome Lid and Completion of Regulatory Particle Assembly. 著者: Robert J Tomko / David W Taylor / Zhuo A Chen / Hong-Wei Wang / Juri Rappsilber / Mark Hochstrasser / ![]() ![]() ![]() ![]() 要旨: Most short-lived eukaryotic proteins are degraded by the proteasome. A proteolytic core particle (CP) capped by regulatory particles (RPs) constitutes the 26S proteasome complex. RP biogenesis ...Most short-lived eukaryotic proteins are degraded by the proteasome. A proteolytic core particle (CP) capped by regulatory particles (RPs) constitutes the 26S proteasome complex. RP biogenesis culminates with the joining of two large subcomplexes, the lid and base. In yeast and mammals, the lid appears to assemble completely before attaching to the base, but how this hierarchical assembly is enforced has remained unclear. Using biochemical reconstitutions, quantitative cross-linking/mass spectrometry, and electron microscopy, we resolve the mechanistic basis for the linkage between lid biogenesis and lid-base joining. Assimilation of the final lid subunit, Rpn12, triggers a large-scale conformational remodeling of the nascent lid that drives RP assembly, in part by relieving steric clash with the base. Surprisingly, this remodeling is triggered by a single Rpn12 α helix. Such assembly-coupled conformational switching is reminiscent of viral particle maturation and may represent a commonly used mechanism to enforce hierarchical assembly in multisubunit complexes. | |||||||||
履歴 |
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構造の表示
ムービー |
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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ダウンロードとリンク
-EMDBアーカイブ
マップデータ | ![]() | 14.7 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 18.8 KB 18.8 KB | 表示 表示 | ![]() |
画像 | ![]() | 149.7 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 219.7 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 218.8 KB | 表示 | |
XML形式データ | ![]() | 5.5 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
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リンク
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Reconstruction of LP2 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 2.8 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
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試料の構成要素
-全体 : Recombinant LP2 (Lid Particle 2)
全体 | 名称: Recombinant LP2 (Lid Particle 2) |
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要素 |
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-超分子 #1000: Recombinant LP2 (Lid Particle 2)
超分子 | 名称: Recombinant LP2 (Lid Particle 2) / タイプ: sample / ID: 1000 / 詳細: The sample was monodisperse. 集合状態: 1 Rpn3: 1 Rpn5: 1 Rpn6: 1 Rpn7: 1 Rpn8: 1 Rpn9: 1 Rpn11: 1 Sem1 Number unique components: 8 |
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分子量 | 理論値: 343 KDa |
-分子 #1: 26S proteasome regulatory subunit RPN3
分子 | 名称: 26S proteasome regulatory subunit RPN3 / タイプ: protein_or_peptide / ID: 1 / Name.synonym: RPN3 / コピー数: 1 / 集合状態: monomer / 組換発現: No |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 64 KDa |
配列 | UniProtKB: 26S proteasome regulatory subunit RPN3 GO: ubiquitin-dependent protein catabolic process, regulation of protein catabolic process, molecular_function, proteasome complex, proteasome regulatory particle, lid subcomplex InterPro: 26S proteasome regulatory subunit, C-terminal, Proteasome component (PCI) domain |
-分子 #2: 26S proteasome regulatory subunit RPN5
分子 | 名称: 26S proteasome regulatory subunit RPN5 / タイプ: protein_or_peptide / ID: 2 / Name.synonym: RPN5 / コピー数: 1 / 集合状態: monomer / 組換発現: No |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 52 KDa |
配列 | UniProtKB: 26S proteasome regulatory subunit RPN5 GO: ubiquitin-dependent protein catabolic process, protein deneddylation => GO:0000338, molecular_function, proteasome complex, proteasome regulatory particle, lid subcomplex InterPro: Proteasome component (PCI) domain, ArsR-like helix-turn-helix domain |
-分子 #3: 26S proteasome regulatory subunit RPN6
分子 | 名称: 26S proteasome regulatory subunit RPN6 / タイプ: protein_or_peptide / ID: 3 / Name.synonym: RPN6 / コピー数: 1 / 集合状態: monomer / 組換発現: No |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 50 KDa |
配列 | UniProtKB: 26S proteasome regulatory subunit RPN6 GO: ubiquitin-dependent protein catabolic process, structural molecule activity, proteasome complex, proteasome regulatory particle, lid subcomplex InterPro: Proteasome component (PCI) domain, INTERPRO: IPR013143 |
-分子 #4: 26S proteasome regulatory subunit RPN7
分子 | 名称: 26S proteasome regulatory subunit RPN7 / タイプ: protein_or_peptide / ID: 4 / Name.synonym: RPN7 / コピー数: 1 / 集合状態: monomer / 組換発現: No |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 49 KDa |
配列 | UniProtKB: 26S proteasome regulatory subunit RPN7 GO: ubiquitin-dependent protein catabolic process, structural molecule activity, proteasome complex, proteasome regulatory particle, lid subcomplex InterPro: 26S proteasome regulatory subunit Rpn7/COP9 signalosome complex subunit 1, Proteasome component (PCI) domain, INTERPRO: IPR013026 |
-分子 #5: 26S proteasome regulatory subunit RPN8
分子 | 名称: 26S proteasome regulatory subunit RPN8 / タイプ: protein_or_peptide / ID: 5 / Name.synonym: RPN8 / コピー数: 1 / 集合状態: monomer / 組換発現: No |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 38 KDa |
配列 | UniProtKB: 26S proteasome regulatory subunit RPN8 GO: ubiquitin-dependent protein catabolic process, molecular_function, proteasome complex, proteasome regulatory particle, lid subcomplex InterPro: JAB1/MPN/MOV34 metalloenzyme domain, Rpn11/EIF3F, C-terminal |
-分子 #6: 26S proteasome regulatory subunit RPN9
分子 | 名称: 26S proteasome regulatory subunit RPN9 / タイプ: protein_or_peptide / ID: 6 / Name.synonym: RPN9 / コピー数: 1 / 集合状態: monomer / 組換発現: No |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 46 KDa |
配列 | UniProtKB: 26S proteasome regulatory subunit RPN9 GO: ubiquitin-dependent protein catabolic process, structural molecule activity, proteasome complex, proteasome regulatory particle, lid subcomplex InterPro: Proteasome component (PCI) domain, ArsR-like helix-turn-helix domain |
-分子 #7: 26S proteasome regulatory subunit RPN11
分子 | 名称: 26S proteasome regulatory subunit RPN11 / タイプ: protein_or_peptide / ID: 7 / Name.synonym: RPN11 / コピー数: 1 / 集合状態: monomer / 組換発現: No |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 34 KDa |
配列 | UniProtKB: Ubiquitin carboxyl-terminal hydrolase RPN11 GO: protein deubiquitination, mitochondrial fission, cysteine-type deubiquitinase activity, proteasome complex, proteasome regulatory particle, lid subcomplex InterPro: JAB1/MPN/MOV34 metalloenzyme domain, Rpn11/EIF3F, C-terminal |
-分子 #8: 26S proteasome regulatory subunit Sem1
分子 | 名称: 26S proteasome regulatory subunit Sem1 / タイプ: protein_or_peptide / ID: 8 / コピー数: 1 / 集合状態: monomer / 組換発現: No |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 10 KDa |
配列 | UniProtKB: 26S proteasome complex subunit SEM1 GO: proteasome complex, proteasome regulatory particle, lid subcomplex, proteasome-mediated ubiquitin-dependent protein catabolic process, proteasome assembly InterPro: DSS1/SEM1 |
-実験情報
-構造解析
手法 | ネガティブ染色法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
濃度 | 0.04 mg/mL |
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緩衝液 | pH: 7.5 詳細: 50 mM Tris, 150 mM NaCl, 5 mM MgCl2, and 10% glycerol |
染色 | タイプ: NEGATIVE 詳細: Sample was negatively stained with six consecutive droplets of 2% uranyl acetate. |
グリッド | 詳細: 400 mesh Cu grid with thin carbon, glow discharged with a sputter coater. |
凍結 | 凍結剤: NONE / 装置: OTHER |
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電子顕微鏡法
顕微鏡 | FEI TECNAI 20 |
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詳細 | Data acquired using Leginon. |
日付 | 2013年4月26日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: GATAN ULTRASCAN 4000 (4k x 4k) 実像数: 600 / 平均電子線量: 30 e/Å2 |
電子線 | 加速電圧: 120 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.2 mm / 最大 デフォーカス(公称値): 1.4 µm / 最小 デフォーカス(公称値): 0.5 µm / 倍率(公称値): 80000 |
試料ステージ | 試料ホルダーモデル: SIDE ENTRY, EUCENTRIC |
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画像解析
CTF補正 | 詳細: Each micrograph |
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最終 再構成 | 想定した対称性 - 点群: C1 (非対称) / アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 16.0 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: EMAN2, SPARX / 使用した粒子像数: 60000 |