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基本情報
登録情報 | ![]() | |||||||||
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タイトル | CryoEM structure of recombinant Trypanosoma cruzi apo proteasome 20S subunit | |||||||||
![]() | Post-processed masked map from Relion used to build model. | |||||||||
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![]() | Proteasome / UNKNOWN FUNCTION | |||||||||
機能・相同性 | ![]() proteasome core complex / proteasome core complex, beta-subunit complex / threonine-type endopeptidase activity / proteasome core complex, alpha-subunit complex / proteasomal protein catabolic process / proteolysis involved in protein catabolic process / ubiquitin-dependent protein catabolic process / proteasome-mediated ubiquitin-dependent protein catabolic process / nucleus / membrane / cytoplasm 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: 2.25 Å | |||||||||
![]() | Rowland P | |||||||||
資金援助 | 1件
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![]() | ![]() タイトル: Pharmacological and structural understanding of the Trypanosoma cruzi proteasome provides key insights for developing site-specific inhibitors. 著者: Thomas C Eadsforth / Leah S Torrie / Paul Rowland / Emma V Edgar / Lorna M MacLean / Christy Paterson / David A Robinson / Sharon M Shepherd / John Thomas / Michael G Thomas / David W Gray / ...著者: Thomas C Eadsforth / Leah S Torrie / Paul Rowland / Emma V Edgar / Lorna M MacLean / Christy Paterson / David A Robinson / Sharon M Shepherd / John Thomas / Michael G Thomas / David W Gray / Vincent L G Postis / Manu De Rycker / ![]() 要旨: The proteasome is considered an excellent drug target for many infectious diseases as well as cancer. Challenges with robust and safe supply of proteasomes from infectious agents, lack of structural ...The proteasome is considered an excellent drug target for many infectious diseases as well as cancer. Challenges with robust and safe supply of proteasomes from infectious agents, lack of structural information, and complex pharmacology due to multiple active sites have hampered progress in the infectious disease space. We recombinantly expressed the proteasome of the protozoan parasite Trypanosoma cruzi, the causative agent of Chagas disease, and demonstrate pharmacological equivalence to the native T. cruzi proteasome. Active-site mutant recombinant proteasomes reveal substrate promiscuity for WT proteasomes, with important implications for assessing pharmacological responses of active-site selective inhibitors. Using these mutant proteasomes, we show that some selective parasite proteasome inhibitors only partially inhibit the chymotrypsin-like activity, including a newly developed 5-(phenoxymethyl)furan-2-carboxamide-based proteasome inhibitor. In spite of partial inhibition, these compounds remain potent inhibitors of intracellular T. cruzi growth. Drug-resistant mutants provide further insights in drug mode-of-inhibition. We also present the high-resolution CryoEM structures of both native and recombinantly-expressed T. cruzi proteasomes which reveal pharmacologically relevant differences in the ligand-binding site compared to the related Leishmania proteasome. Furthermore, we show that the trypanosomatid β4/β5 selectivity pocket is not present in the proteasome structures of other protozoan parasites. This work highlights the need, and provides approaches, to precisely assess proteasome substrate selectivity and pharmacology. It enables structure-guided drug discovery for this promising Chagas disease drug target, provides a new chemical starting point for drug discovery, and paves the road for development of robust proteasome drug discovery programmes for other eukaryotic infectious diseases. | |||||||||
履歴 |
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構造の表示
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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 | ![]() | 40.1 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 30.4 KB 30.4 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 17 KB | 表示 | ![]() |
画像 | ![]() | 202.9 KB | ||
Filedesc metadata | ![]() | 8.1 KB | ||
その他 | ![]() ![]() | 336.9 MB 335.9 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 959.3 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 958.8 KB | 表示 | |
XML形式データ | ![]() | 24.7 KB | 表示 | |
CIF形式データ | ![]() | 32.7 KB | 表示 | |
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 | ![]() 9f9pMC ![]() 9f9tC 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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注釈 | Post-processed masked map from Relion used to build model. | ||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 0.66 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
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試料の構成要素
+全体 : Proteasome 20S
+超分子 #1: Proteasome 20S
+分子 #1: Proteasome subunit alpha type
+分子 #2: Proteasome subunit alpha type
+分子 #3: Proteasome subunit alpha type
+分子 #4: Proteasome subunit alpha type
+分子 #5: Proteasome subunit alpha type
+分子 #6: Proteasome subunit alpha type
+分子 #7: Putative proteasome alpha 7 subunit
+分子 #8: Proteasome subunit beta
+分子 #9: Proteasome subunit beta
+分子 #10: Putative proteasome beta 3 subunit
+分子 #11: Proteasome subunit beta
+分子 #12: Proteasome subunit beta
+分子 #13: Proteasome subunit beta
+分子 #14: Proteasome subunit beta
+分子 #15: water
-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: 7.5 |
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凍結 | 凍結剤: ETHANE |
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電子顕微鏡法
顕微鏡 | FEI TITAN 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: 1.05 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: OTHER / 撮影モード: OTHER / 最大 デフォーカス(公称値): 2.0 µm / 最小 デフォーカス(公称値): 1.0 µm |
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: 2.25 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 92013 |
初期 角度割当 | タイプ: OTHER |
最終 角度割当 | タイプ: OTHER |