+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 5khr | ||||||
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タイトル | Model of human Anaphase-promoting complex/Cyclosome complex (APC15 deletion mutant) in complex with the E2 UBE2C/UBCH10 poised for ubiquitin ligation to substrate (APC/C-CDC20-substrate-UBE2C) | ||||||
要素 |
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キーワード | CELL CYCLE / ubiquitination / protein complex / mitosis | ||||||
機能・相同性 | 機能・相同性情報 protein localization to septin ring / mitotic morphogenesis checkpoint signaling / metaphase/anaphase transition of cell cycle / metaphase/anaphase transition of meiosis I / cellular bud neck septin ring / Inhibition of the proteolytic activity of APC/C required for the onset of anaphase by mitotic spindle checkpoint components / mitotic checkpoint complex / positive regulation of anaphase-promoting complex-dependent catabolic process / positive regulation of exit from mitosis / regulation of meiotic nuclear division ...protein localization to septin ring / mitotic morphogenesis checkpoint signaling / metaphase/anaphase transition of cell cycle / metaphase/anaphase transition of meiosis I / cellular bud neck septin ring / Inhibition of the proteolytic activity of APC/C required for the onset of anaphase by mitotic spindle checkpoint components / mitotic checkpoint complex / positive regulation of anaphase-promoting complex-dependent catabolic process / positive regulation of exit from mitosis / regulation of meiotic nuclear division / free ubiquitin chain polymerization / positive regulation of synapse maturation / Conversion from APC/C:Cdc20 to APC/C:Cdh1 in late anaphase / protein branched polyubiquitination / Inactivation of APC/C via direct inhibition of the APC/C complex / APC/C:Cdc20 mediated degradation of mitotic proteins / Phosphorylation of Emi1 / anaphase-promoting complex / Aberrant regulation of mitotic exit in cancer due to RB1 defects / regulation of meiotic cell cycle / anaphase-promoting complex-dependent catabolic process / metaphase/anaphase transition of mitotic cell cycle / positive regulation of synaptic plasticity / (E3-independent) E2 ubiquitin-conjugating enzyme / regulation of exit from mitosis / anaphase-promoting complex binding / Phosphorylation of the APC/C / cellular bud neck / positive regulation of mitotic metaphase/anaphase transition / ubiquitin ligase activator activity / positive regulation of ubiquitin protein ligase activity / protein K11-linked ubiquitination / enzyme-substrate adaptor activity / positive regulation of dendrite morphogenesis / regulation of mitotic metaphase/anaphase transition / ubiquitin-ubiquitin ligase activity / exit from mitosis / mitotic sister chromatid cohesion / mitotic metaphase chromosome alignment / E2 ubiquitin-conjugating enzyme / mitotic spindle assembly checkpoint signaling / ubiquitin-like protein ligase binding / Regulation of APC/C activators between G1/S and early anaphase / ubiquitin conjugating enzyme activity / cullin family protein binding / Transcriptional Regulation by VENTX / mitotic spindle assembly / ubiquitin-like ligase-substrate adaptor activity / positive regulation of axon extension / protein K48-linked ubiquitination / heterochromatin / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / ubiquitin ligase complex / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / regulation of mitotic cell cycle / Resolution of Sister Chromatid Cohesion / APC/C:Cdc20 mediated degradation of Cyclin B / APC-Cdc20 mediated degradation of Nek2A / nuclear periphery / Synthesis of active ubiquitin: roles of E1 and E2 enzymes / Autodegradation of Cdh1 by Cdh1:APC/C / APC/C:Cdc20 mediated degradation of Securin / SCF-beta-TrCP mediated degradation of Emi1 / RHO GTPases Activate Formins / Assembly of the pre-replicative complex / Cdc20:Phospho-APC/C mediated degradation of Cyclin A / brain development / APC/C:Cdh1 mediated degradation of Cdc20 and other APC/C:Cdh1 targeted proteins in late mitosis/early G1 / protein catabolic process / CDK-mediated phosphorylation and removal of Cdc6 / mitotic spindle / kinetochore / spindle pole / spindle / protein polyubiquitination / ubiquitin-protein transferase activity / Separation of Sister Chromatids / G2/M transition of mitotic cell cycle / microtubule cytoskeleton / ubiquitin protein ligase activity / Antigen processing: Ubiquitination & Proteasome degradation / mitotic cell cycle / nervous system development / Senescence-Associated Secretory Phenotype (SASP) / ubiquitin-dependent protein catabolic process / protein phosphatase binding / proteasome-mediated ubiquitin-dependent protein catabolic process / molecular adaptor activity / cell differentiation / non-specific serine/threonine protein kinase / regulation of cell cycle / protein kinase activity / Ub-specific processing proteases / protein ubiquitination / cell division / negative regulation of gene expression / protein serine kinase activity / protein serine/threonine kinase activity / centrosome 類似検索 - 分子機能 | ||||||
生物種 | Homo sapiens (ヒト) Saccharomyces cerevisiae (パン酵母) | ||||||
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: 6.1 Å | ||||||
データ登録者 | VanderLinden, R. / Yamaguchi, M. / Dube, P. / Haselbach, D. / Stark, H. / Schulman, B.A. | ||||||
引用 | ジャーナル: Mol Cell / 年: 2016 タイトル: Cryo-EM of Mitotic Checkpoint Complex-Bound APC/C Reveals Reciprocal and Conformational Regulation of Ubiquitin Ligation. 著者: Masaya Yamaguchi / Ryan VanderLinden / Florian Weissmann / Renping Qiao / Prakash Dube / Nicholas G Brown / David Haselbach / Wei Zhang / Sachdev S Sidhu / Jan-Michael Peters / Holger Stark / ...著者: Masaya Yamaguchi / Ryan VanderLinden / Florian Weissmann / Renping Qiao / Prakash Dube / Nicholas G Brown / David Haselbach / Wei Zhang / Sachdev S Sidhu / Jan-Michael Peters / Holger Stark / Brenda A Schulman / 要旨: The mitotic checkpoint complex (MCC) coordinates proper chromosome biorientation on the spindle with ubiquitination activities of CDC20-activated anaphase-promoting complex/cyclosome (APC/C(CDC20)). ...The mitotic checkpoint complex (MCC) coordinates proper chromosome biorientation on the spindle with ubiquitination activities of CDC20-activated anaphase-promoting complex/cyclosome (APC/C(CDC20)). APC/C(CDC20) and two E2s, UBE2C and UBE2S, catalyze ubiquitination through distinct architectures for linking ubiquitin (UB) to substrates and elongating polyUB chains, respectively. MCC, which contains a second molecule of CDC20, blocks APC/C(CDC20)-UBE2C-dependent ubiquitination of Securin and Cyclins, while differentially determining or inhibiting CDC20 ubiquitination to regulate spindle surveillance, checkpoint activation, and checkpoint termination. Here electron microscopy reveals conformational variation of APC/C(CDC20)-MCC underlying this multifaceted regulation. MCC binds APC/C-bound CDC20 to inhibit substrate access. However, rotation about the CDC20-MCC assembly and conformational variability of APC/C modulate UBE2C-catalyzed ubiquitination of MCC's CDC20 molecule. Access of UBE2C is limiting for subsequent polyubiquitination by UBE2S. We propose that conformational dynamics of APC/C(CDC20)-MCC modulate E2 activation and determine distinctive ubiquitination activities as part of a response mechanism ensuring accurate sister chromatid segregation. | ||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | 分子: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 5khr.cif.gz | 354.6 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb5khr.ent.gz | 196.1 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 5khr.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 | 5khr_validation.pdf.gz | 981.5 KB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 5khr_full_validation.pdf.gz | 981.6 KB | 表示 | |
XML形式データ | 5khr_validation.xml.gz | 86.3 KB | 表示 | |
CIF形式データ | 5khr_validation.cif.gz | 136.5 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/kh/5khr ftp://data.pdbj.org/pub/pdb/validation_reports/kh/5khr | HTTPS FTP |
-関連構造データ
関連構造データ | 4025MC 4021C 4022C 4023C 4024C 4026C 4027C 4028C 5khuC 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 (文献) |
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類似構造データ |
-リンク
-集合体
登録構造単位 |
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-要素
-Anaphase-promoting complex subunit ... , 10種, 12分子 ABEGWILMNOXY
#1: タンパク質 | 分子量: 216725.469 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: ANAPC1, TSG24 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q9H1A4 | ||||||||||||
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#2: タンパク質 | 分子量: 9854.647 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: ANAPC11, HSPC214 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q9NYG5 | ||||||||||||
#4: タンパク質 | 分子量: 11677.995 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: ANAPC16, C10orf104, CENP-27 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q96DE5 | ||||||||||||
#6: タンパク質 | 分子量: 9793.999 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: CDC26, ANAPC12, C9orf17 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q8NHZ8 #7: タンパク質 | | 分子量: 93207.328 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: Contains the remnant of a C-terminal linker and HRV14 3C protease site. 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: ANAPC4, APC4 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q9UJX5 #9: タンパク質 | | 分子量: 21282.143 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: ANAPC10, APC10 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q9UM13 #10: タンパク質 | | 分子量: 8528.309 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: ANAPC13 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q9BS18 #11: タンパク質 | | 分子量: 94023.055 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: ANAPC2, APC2, KIAA1406 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q9UJX6 #12: タンパク質 | | 分子量: 85249.812 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: ANAPC5, APC5 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q9UJX4 #16: タンパク質 | 分子量: 63204.020 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: ANAPC7, APC7 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q9UJX3 |
-Cell division cycle protein ... , 4種, 7分子 CPFHJKR
#3: タンパク質 | 分子量: 68921.031 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: CDC23, ANAPC8 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q9UJX2 #5: タンパク質 | 分子量: 91973.125 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: CDC27, ANAPC3, D0S1430E, D17S978E / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: P30260 #8: タンパク質 | 分子量: 71747.516 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: CDC16, ANAPC6 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q13042 #14: タンパク質 | | 分子量: 54796.508 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: CDC20 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q12834 |
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-タンパク質 / タンパク質・ペプチド , 2種, 2分子 QS
#13: タンパク質 | 分子量: 20748.438 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: Contains the remnant of a C-terminal linker and HRV14 3C protease site. 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: UBE2C, UBCH10 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) 参照: UniProt: O00762, E2 ubiquitin-conjugating enzyme, (E3-independent) E2 ubiquitin-conjugating enzyme |
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#15: タンパク質・ペプチド | 分子量: 3783.135 Da / 分子数: 1 / 由来タイプ: 合成 詳細: Contains azidohomoalanine in place of K788 and is cross-linked to UBE2C and UB. 由来: (合成) Saccharomyces cerevisiae (パン酵母) / 参照: UniProt: P34244*PLUS |
-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 |
-試料調製
構成要素 | 名称: Multi-protein complex of APC/C with coactivator CDC20 and cross-linked UBE2C-UB-Substrate complex mimicking ubiquitination タイプ: COMPLEX / Entity ID: all / 由来: MULTIPLE SOURCES |
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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 |
-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: SPOT SCAN |
電子レンズ | モード: BRIGHT FIELD |
撮影 | 電子線照射量: 40 e/Å2 フィルム・検出器のモデル: FEI FALCON II (4k x 4k) |
-解析
CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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3次元再構成 | 解像度: 6.1 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 222697 / 対称性のタイプ: POINT |