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基本情報
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タイトル | Human proteasome in resting state conformation bound to TXNL1 in Forward conformation | |||||||||
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![]() | 26S Proteasome / Nub1 / Fat10 / MOTOR PROTEIN / HYDROLASE-PROTEIN BINDING complex | |||||||||
機能・相同性 | ![]() regulation of ubiquitin-dependent protein catabolic process / positive regulation of inclusion body assembly / disulfide oxidoreductase activity / thyrotropin-releasing hormone receptor binding / modulation by host of viral transcription / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / cytosolic proteasome complex / 加水分解酵素; プロテアーゼ; ペプチド結合加水分解酵素; オメガペプチターゼ / proteasome accessory complex ...regulation of ubiquitin-dependent protein catabolic process / positive regulation of inclusion body assembly / disulfide oxidoreductase activity / thyrotropin-releasing hormone receptor binding / modulation by host of viral transcription / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / cytosolic proteasome complex / 加水分解酵素; プロテアーゼ; ペプチド結合加水分解酵素; オメガペプチターゼ / proteasome accessory complex / integrator complex / purine ribonucleoside triphosphate binding / meiosis I / proteasome regulatory particle / positive regulation of proteasomal protein catabolic process / proteasome-activating activity / proteasome regulatory particle, lid subcomplex / proteasome regulatory particle, base subcomplex / metal-dependent deubiquitinase activity / negative regulation of programmed cell death / protein K63-linked deubiquitination / RHOBTB1 GTPase cycle / Regulation of ornithine decarboxylase (ODC) / RND1 GTPase cycle / RND2 GTPase cycle / Proteasome assembly / RND3 GTPase cycle / Homologous DNA Pairing and Strand Exchange / Defective homologous recombination repair (HRR) due to BRCA1 loss of function / Defective HDR through Homologous Recombination Repair (HRR) due to PALB2 loss of BRCA1 binding function / Defective HDR through Homologous Recombination Repair (HRR) due to PALB2 loss of BRCA2/RAD51/RAD51C binding function / Cross-presentation of soluble exogenous antigens (endosomes) / Resolution of D-loop Structures through Synthesis-Dependent Strand Annealing (SDSA) / proteasome core complex / Resolution of D-loop Structures through Holliday Junction Intermediates / Somitogenesis / K63-linked deubiquitinase activity / RHOV GTPase cycle / Impaired BRCA2 binding to RAD51 / proteasome binding / transcription factor binding / regulation of protein catabolic process / myofibril / proteasome storage granule / RHOU GTPase cycle / Presynaptic phase of homologous DNA pairing and strand exchange / protein-disulfide reductase activity / response to tumor necrosis factor / general transcription initiation factor binding / response to type II interferon / blastocyst development / polyubiquitin modification-dependent protein binding / immune system process / positive regulation of RNA polymerase II transcription preinitiation complex assembly / protein deubiquitination / endopeptidase activator activity / NF-kappaB binding / proteasome assembly / proteasome core complex, alpha-subunit complex / mRNA export from nucleus / RHOBTB2 GTPase cycle / SARS-CoV-1 targets host intracellular signalling and regulatory pathways / inclusion body / enzyme regulator activity / ERAD pathway / regulation of proteasomal protein catabolic process / proteasome complex / proteolysis involved in protein catabolic process / sarcomere / Regulation of activated PAK-2p34 by proteasome mediated degradation / Autodegradation of Cdh1 by Cdh1:APC/C / APC/C:Cdc20 mediated degradation of Securin / N-glycan trimming in the ER and Calnexin/Calreticulin cycle / Asymmetric localization of PCP proteins / Ubiquitin-dependent degradation of Cyclin D / SCF-beta-TrCP mediated degradation of Emi1 / NIK-->noncanonical NF-kB signaling / stem cell differentiation / TNFR2 non-canonical NF-kB pathway / AUF1 (hnRNP D0) binds and destabilizes mRNA / Vpu mediated degradation of CD4 / Assembly of the pre-replicative complex / Ubiquitin Mediated Degradation of Phosphorylated Cdc25A / Degradation of DVL / Cdc20:Phospho-APC/C mediated degradation of Cyclin A / Dectin-1 mediated noncanonical NF-kB signaling / lipopolysaccharide binding / Degradation of AXIN / Hh mutants are degraded by ERAD / negative regulation of inflammatory response to antigenic stimulus / P-body / Activation of NF-kappaB in B cells / Degradation of GLI1 by the proteasome / Hedgehog ligand biogenesis / G2/M Checkpoints / Defective CFTR causes cystic fibrosis / GSK3B and BTRC:CUL1-mediated-degradation of NFE2L2 / Autodegradation of the E3 ubiquitin ligase COP1 / Negative regulation of NOTCH4 signaling / Vif-mediated degradation of APOBEC3G 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: 2.87 Å | |||||||||
![]() | Arkinson C / Gee CL / Martin A | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Structural landscape of AAA+ ATPase motor states in the substrate-degrading human 26S proteasome reveals conformation-specific binding of TXNL1. 著者: Connor Arkinson / Christine L Gee / Zeyuan Zhang / Ken C Dong / Andreas Martin / ![]() 要旨: The 26S proteasome targets many cellular proteins for degradation during general homeostasis, protein quality control, and the regulation of vital processes. A broad range of proteasome-interacting ...The 26S proteasome targets many cellular proteins for degradation during general homeostasis, protein quality control, and the regulation of vital processes. A broad range of proteasome-interacting cofactors thereby modulates these functions and aids in substrate degradation. Here, we solved several high-resolution structures of the redox active cofactor TXNL1 bound to the human 26S proteasome at saturating and sub-stoichiometric concentrations by time resolved cryo-EM. We identified distinct binding modes of TXNL1 that depend on the proteasome conformational and ATPase motor states. Together with biophysical and biochemical experiments, our structural studies reveal that the resting-state proteasome prior to substrate engagement with the ATPase motor binds TXNL1 with low affinity and in variable positions on top of the Rpn11 deubiquitinase. In contrast, the actively degrading proteasome shows additional interactions leading to high-affinity TXNL1 binding, whereby TXNL1's C-terminal tail covers the catalytic groove of the Rpn11 deubiquitinase and coordinates the active-site Zn. Furthermore, these cryo-EM structures of the degrading proteasome capture the ATPase hexamer in all registers of spiral-staircase arrangements and thus visualize the complete ATP-hydrolysis cycle of the AAA+ motor, indicating temporally asymmetric hydrolysis and conformational changes in bursts during mechanical substrate unfolding and translocation. Remarkably, we catch the proteasome in the act of unfolding the beta-barrel mEos3.2 substrate while the ATPase hexamer is in a particular spiral staircase register. Our findings challenge current models for protein translocation through hexameric AAA+ motors and reveal how the proteasome uses its distinct but broad range of conformational states to coordinate cofactor binding and substrate processing. | |||||||||
履歴 |
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構造の表示
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 | ![]() | 75.8 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 48.1 KB 48.1 KB | 表示 表示 | ![]() |
画像 | ![]() | 121 KB | ||
Filedesc metadata | ![]() | 13.5 KB | ||
その他 | ![]() ![]() | 139.2 MB 139.2 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 9e8iMC ![]() 9e8gC ![]() 9e8hC ![]() 9e8jC ![]() 9e8kC ![]() 9e8lC ![]() 9e8nC ![]() 9e8oC ![]() 9e8qC M: このマップから作成された原子モデル 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 ![]() |
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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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投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.048 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-ハーフマップ: #1
ファイル | emd_47721_half_map_1.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: #2
ファイル | emd_47721_half_map_2.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
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試料の構成要素
+全体 : Human 26S proteasome complexed with Nub1, Fat10 and TXNL1
+超分子 #1: Human 26S proteasome complexed with Nub1, Fat10 and TXNL1
+分子 #1: 26S proteasome regulatory subunit 7
+分子 #2: 26S proteasome regulatory subunit 4
+分子 #3: 26S protease regulatory subunit 8
+分子 #4: 26S proteasome regulatory subunit 6B
+分子 #5: 26S protease regulatory subunit 10B
+分子 #6: 26S proteasome regulatory subunit 6A
+分子 #7: Proteasome subunit alpha type-6
+分子 #8: Proteasome subunit alpha type-2
+分子 #9: Proteasome subunit alpha type-4
+分子 #10: Proteasome subunit alpha type-7
+分子 #11: Proteasome subunit alpha type-5
+分子 #12: Proteasome subunit alpha type-1
+分子 #13: Proteasome subunit alpha type-3
+分子 #14: 26S proteasome non-ATPase regulatory subunit 1
+分子 #15: 26S proteasome non-ATPase regulatory subunit 3
+分子 #16: 26S proteasome non-ATPase regulatory subunit 12
+分子 #17: 26S proteasome non-ATPase regulatory subunit 11
+分子 #18: 26S proteasome non-ATPase regulatory subunit 6
+分子 #19: 26S proteasome non-ATPase regulatory subunit 7
+分子 #20: 26S proteasome non-ATPase regulatory subunit 13
+分子 #21: 26S proteasome non-ATPase regulatory subunit 4
+分子 #22: 26S proteasome non-ATPase regulatory subunit 14
+分子 #23: 26S proteasome non-ATPase regulatory subunit 8
+分子 #24: 26S proteasome complex subunit SEM1
+分子 #25: 26S proteasome non-ATPase regulatory subunit 2
+分子 #26: Isoform 2 of NEDD8 ultimate buster 1
+分子 #27: Thioredoxin-like protein 1
+分子 #28: ADENOSINE-5'-TRIPHOSPHATE
+分子 #29: MAGNESIUM ION
+分子 #30: ADENOSINE-5'-DIPHOSPHATE
+分子 #31: ZINC ION
-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
緩衝液 | pH: 7.4 詳細: 30 mM HEPES pH7.4, 25 mM NaCl, 25 mM KCl, 3% (v/v) glycerol, 5 mM MgCl2 2 mM ATP and 0.5 mM TCEP. |
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凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 285 K / 装置: FEI VITROBOT MARK IV |
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電子顕微鏡法
顕微鏡 | TFS KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 平均電子線量: 50.0 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 / 最大 デフォーカス(公称値): 1.7 µm / 最小 デフォーカス(公称値): 0.5 µm |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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画像解析
初期モデル | モデルのタイプ: NONE |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 2.87 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 62431 |
初期 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD |