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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: EMDB / ID: EMD-2925 | |||||||||
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タイトル | Cryo-EM structure of the human APC/C-Cdh1-Hsl1-UbcH10-Ub complex. | |||||||||
![]() | Cryo-EM structure of the human APC/C-Cdh1-Hsl1-UbcH10-Ub complex. | |||||||||
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![]() | Ubiquitination / Cell cycle. | |||||||||
機能・相同性 | ![]() negative regulation of mitotic spindle pole body separation / Conversion from APC/C:Cdc20 to APC/C:Cdh1 in late anaphase / positive regulation of mitotic actomyosin contractile ring contraction / deactivation of mitotic spindle assembly checkpoint / positive regulation of anaphase-promoting complex-dependent catabolic process / positive regulation of exit from mitosis / Antigen processing: Ubiquitination & Proteasome degradation / free ubiquitin chain polymerization / Conversion from APC/C:Cdc20 to APC/C:Cdh1 in late anaphase / regulation of mitotic cell cycle spindle assembly checkpoint ...negative regulation of mitotic spindle pole body separation / Conversion from APC/C:Cdc20 to APC/C:Cdh1 in late anaphase / positive regulation of mitotic actomyosin contractile ring contraction / deactivation of mitotic spindle assembly checkpoint / positive regulation of anaphase-promoting complex-dependent catabolic process / positive regulation of exit from mitosis / Antigen processing: Ubiquitination & Proteasome degradation / free ubiquitin chain polymerization / Conversion from APC/C:Cdc20 to APC/C:Cdh1 in late anaphase / regulation of mitotic cell cycle spindle assembly checkpoint / positive regulation of synapse maturation / Inactivation of APC/C via direct inhibition of the APC/C complex / APC/C:Cdc20 mediated degradation of mitotic proteins / anaphase-promoting complex / Aberrant regulation of mitotic exit in cancer due to RB1 defects / regulation of meiotic cell cycle / metaphase/anaphase transition of mitotic cell cycle / anaphase-promoting complex-dependent catabolic process / (E3-independent) E2 ubiquitin-conjugating enzyme / positive regulation of synaptic plasticity / regulation of exit from mitosis / anaphase-promoting complex binding / Phosphorylation of the APC/C / positive regulation of mitotic metaphase/anaphase transition / positive regulation of ubiquitin protein ligase activity / ubiquitin ligase activator activity / Regulation of APC/C activators between G1/S and early anaphase / positive regulation of ubiquitin-dependent protein catabolic process / protein K11-linked ubiquitination / enzyme-substrate adaptor activity / positive regulation of dendrite morphogenesis / regulation of mitotic metaphase/anaphase transition / cell cycle / exit from mitosis / ubiquitin-ubiquitin ligase activity / mitotic metaphase chromosome alignment / E2 ubiquitin-conjugating enzyme / ubiquitin-like protein ligase binding / regulation of mitotic cell cycle / cullin family protein binding / ubiquitin conjugating enzyme activity / Antigen processing: Ubiquitination & Proteasome degradation / cyclin binding / Transcriptional Regulation by VENTX / positive regulation of axon extension / protein K48-linked ubiquitination / heterochromatin / ubiquitin ligase complex / APC/C:Cdc20 mediated degradation of Cyclin B / APC-Cdc20 mediated degradation of Nek2A / Synthesis of active ubiquitin: roles of E1 and E2 enzymes / nuclear periphery / Autodegradation of Cdh1 by Cdh1:APC/C / APC/C:Cdc20 mediated degradation of Securin / Assembly of the pre-replicative complex / CDK-mediated phosphorylation and removal of Cdc6 / Cdc20:Phospho-APC/C mediated degradation of Cyclin A / APC/C:Cdh1 mediated degradation of Cdc20 and other APC/C:Cdh1 targeted proteins in late mitosis/early G1 / brain development / mitotic spindle / kinetochore / spindle / protein polyubiquitination / positive regulation of protein catabolic process / Separation of Sister Chromatids / ubiquitin-protein transferase activity / microtubule cytoskeleton / ubiquitin protein ligase activity / Antigen processing: Ubiquitination & Proteasome degradation / Senescence-Associated Secretory Phenotype (SASP) / mitotic cell cycle / nervous system development / protein phosphatase binding / ubiquitin-dependent protein catabolic process / cell differentiation / molecular adaptor activity / protein ubiquitination / cell division / negative regulation of gene expression / centrosome / ubiquitin protein ligase binding / nucleolus / zinc ion binding / nucleoplasm / ATP binding / nucleus / plasma membrane / cytoplasm / cytosol 類似検索 - 分子機能 | |||||||||
生物種 | ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 5.7 Å | |||||||||
![]() | Chang L / Zhang Z / Yang J / McLaughlin SH / Barford D | |||||||||
![]() | ![]() タイトル: Atomic structure of the APC/C and its mechanism of protein ubiquitination. 著者: Leifu Chang / Ziguo Zhang / Jing Yang / Stephen H McLaughlin / David Barford / ![]() 要旨: The anaphase-promoting complex (APC/C) is a multimeric RING E3 ubiquitin ligase that controls chromosome segregation and mitotic exit. Its regulation by coactivator subunits, phosphorylation, the ...The anaphase-promoting complex (APC/C) is a multimeric RING E3 ubiquitin ligase that controls chromosome segregation and mitotic exit. Its regulation by coactivator subunits, phosphorylation, the mitotic checkpoint complex and interphase early mitotic inhibitor 1 (Emi1) ensures the correct order and timing of distinct cell-cycle transitions. Here we use cryo-electron microscopy to determine atomic structures of APC/C-coactivator complexes with either Emi1 or a UbcH10-ubiquitin conjugate. These structures define the architecture of all APC/C subunits, the position of the catalytic module and explain how Emi1 mediates inhibition of the two E2s UbcH10 and Ube2S. Definition of Cdh1 interactions with the APC/C indicates how they are antagonized by Cdh1 phosphorylation. The structure of the APC/C with UbcH10-ubiquitin reveals insights into the initiating ubiquitination reaction. Our results provide a quantitative framework for the design of future experiments to investigate APC/C functions in vivo. | |||||||||
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
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マップデータ | ![]() | 4.3 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 18.7 KB 18.7 KB | 表示 表示 | ![]() |
画像 | ![]() | 118.8 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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リンク
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「今月の分子」の関連する項目 |
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ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Cryo-EM structure of the human APC/C-Cdh1-Hsl1-UbcH10-Ub complex. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.77 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 human APC/C-Cdh1-Hsl1-UbcH10LR-Ub complex
+超分子 #1000: Recombinant human APC/C-Cdh1-Hsl1-UbcH10LR-Ub complex
+分子 #1: Apc1
+分子 #2: Apc2
+分子 #3: Apc3
+分子 #4: Apc4
+分子 #5: Apc5
+分子 #6: Apc6
+分子 #7: Apc7
+分子 #8: Apc8
+分子 #9: Apc10
+分子 #10: Apc11
+分子 #11: Apc12
+分子 #12: Apc13
+分子 #13: Apc15
+分子 #14: Apc16
+分子 #15: Cdh1
+分子 #16: Hsl1
+分子 #17: E2-ubiquitin conjugate
+分子 #18: UbcH10-ubiquitin
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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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 | 0.2 mg/mL |
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緩衝液 | pH: 8 / 詳細: 20 mM HEPES, 150 mM NaCl, 1 mM DTT |
グリッド | 詳細: R2/2 Quantifoil grid with thin carbon support. |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / 装置: FEI VITROBOT MARK III / 手法: Blot for 5 seconds before plunging |
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電子顕微鏡法
顕微鏡 | FEI POLARA 300 |
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日付 | 2014年7月12日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: FEI FALCON II (4k x 4k) 平均電子線量: 16 e/Å2 |
電子線 | 加速電圧: 300 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: 2.0 mm / 最大 デフォーカス(公称値): 4.0 µm / 最小 デフォーカス(公称値): 2.0 µm / 倍率(公称値): 59000 |
試料ステージ | 試料ホルダーモデル: OTHER |
実験機器 | ![]() モデル: Tecnai Polara / 画像提供: FEI Company |
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画像解析
詳細 | The particles were selected using an automatic selection program |
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CTF補正 | 詳細: Each micrograph |
最終 再構成 | 想定した対称性 - 点群: C1 (非対称) / 解像度のタイプ: BY AUTHOR / 解像度: 5.7 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: RELION / 使用した粒子像数: 19939 |