+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-23669 | |||||||||
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タイトル | A. baumannii Ribosome-Eravacycline complex: P-site tRNA 70S | |||||||||
マップデータ | Combined density modified map of Acinetobacter baumannii 70S-Eravacycline with P-site tRNA | |||||||||
試料 |
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キーワード | Acinetobacter baumannii / ribosome / eravacycline / RIBOSOME-ANTIBIOTIC complex | |||||||||
機能・相同性 | 機能・相同性情報 ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation ...ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / ribonucleoprotein complex / translation / mRNA binding / RNA binding / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | Acinetobacter baumannii (strain AB0057) (バクテリア) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 2.66 Å | |||||||||
データ登録者 | Morgan CE / Yu EW | |||||||||
資金援助 | 米国, 1件
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引用 | ジャーナル: mBio / 年: 2021 タイトル: Cryo-EM Determination of Eravacycline-Bound Structures of the Ribosome and the Multidrug Efflux Pump AdeJ of Acinetobacter baumannii. 著者: Zhemin Zhang / Christopher E Morgan / Robert A Bonomo / Edward W Yu / 要旨: Antibiotic-resistant strains of the Gram-negative pathogen Acinetobacter baumannii have emerged as a significant global health threat. One successful therapeutic option to treat bacterial infections ...Antibiotic-resistant strains of the Gram-negative pathogen Acinetobacter baumannii have emerged as a significant global health threat. One successful therapeutic option to treat bacterial infections has been to target the bacterial ribosome. However, in many cases, multidrug efflux pumps within the bacterium recognize and extrude these clinically important antibiotics designed to inhibit the protein synthesis function of the bacterial ribosome. Thus, multidrug efflux within A. baumannii and other highly drug-resistant strains is a major cause of failure of drug-based treatments of infectious diseases. We here report the first structures of the cinetobacter rug fflux (Ade)J pump in the presence of the antibiotic eravacycline, using single-particle cryo-electron microscopy (cryo-EM). We also describe cryo-EM structures of the eravacycline-bound forms of the A. baumannii ribosome, including the 70S, 50S, and 30S forms. Our data indicate that the AdeJ pump primarily uses hydrophobic interactions to bind eravacycline, while the 70S ribosome utilizes electrostatic interactions to bind this drug. Our work here highlights how an antibiotic can bind multiple bacterial targets through different mechanisms and potentially enables drug optimization by taking advantage of these different modes of ligand binding. Acinetobacter baumannii has developed into a highly antibiotic-resistant Gram-negative pathogen. The prevalent AdeJ multidrug efflux pump mediates resistance to different classes of antibiotics known to inhibit the function of the 70S ribosome. Here, we report the first structures of the A. baumannii AdeJ pump, both in the absence and presence of eravacycline. We also describe structures of the A. baumannii ribosome bound by this antibiotic. Our results indicate that AdeJ and the ribosome use very distinct binding modes for drug recognition. Our work will ultimately enable structure-based drug discovery to combat antibiotic-resistant A. baumannii infection. | |||||||||
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 | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_23669.map.gz | 67.5 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-23669-v30.xml emd-23669.xml | 73.9 KB 73.9 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_23669.png | 91 KB | ||
Filedesc metadata | emd-23669.cif.gz | 13.5 KB | ||
その他 | emd_23669_additional_1.map.gz emd_23669_additional_2.map.gz emd_23669_additional_3.map.gz | 90.7 MB 47 MB 24.4 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-23669 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-23669 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_23669_validation.pdf.gz | 445.1 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_23669_full_validation.pdf.gz | 444.6 KB | 表示 | |
XML形式データ | emd_23669_validation.xml.gz | 7.9 KB | 表示 | |
CIF形式データ | emd_23669_validation.cif.gz | 9.1 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23669 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23669 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_23669.map.gz / 形式: CCP4 / 大きさ: 512 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Combined density modified map of Acinetobacter baumannii 70S-Eravacycline with P-site tRNA | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 0.848 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
-追加マップ: Focus-refined and density modified map of 50S from...
ファイル | emd_23669_additional_1.map | ||||||||||||
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注釈 | Focus-refined and density modified map of 50S from Acinetobacter baumannii 70S-Eravacycline with P-site tRNA | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Focus-refined and density modified map of 30S core...
ファイル | emd_23669_additional_2.map | ||||||||||||
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注釈 | Focus-refined and density modified map of 30S core from Acinetobacter baumannii 70S-Eravacycline with P-site tRNA | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Focus-refined and density modified map of 30S head...
ファイル | emd_23669_additional_3.map | ||||||||||||
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注釈 | Focus-refined and density modified map of 30S head from Acinetobacter baumannii 70S-Eravacycline with P-site tRNA | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
+全体 : A baumannii 70S ribosome with P-site tRNA in complex with Eravacycline
+超分子 #1: A baumannii 70S ribosome with P-site tRNA in complex with Eravacycline
+分子 #1: 50S ribosomal protein L33
+分子 #2: 50S ribosomal protein L34
+分子 #3: 50S ribosomal protein L35
+分子 #4: 50S ribosomal protein L36
+分子 #7: 50S ribosomal protein L2
+分子 #8: 50S ribosomal protein L3
+分子 #9: 50S ribosomal protein L4
+分子 #10: 50S ribosomal protein L5
+分子 #11: 50S ribosomal protein L6
+分子 #12: 50S ribosomal protein L9
+分子 #13: 50S ribosomal protein L13
+分子 #14: 50S ribosomal protein L14
+分子 #15: 50S ribosomal protein L15
+分子 #16: 50S ribosomal protein L16
+分子 #17: 50S ribosomal protein L17
+分子 #18: 50S ribosomal protein L18
+分子 #19: 50S ribosomal protein L19
+分子 #20: 50S ribosomal protein L20
+分子 #21: 50S ribosomal protein L21
+分子 #22: 50S ribosomal protein L22
+分子 #23: 50S ribosomal protein L23
+分子 #24: 50S ribosomal protein L24
+分子 #25: 50S ribosomal protein L25
+分子 #26: 50S ribosomal protein L27
+分子 #27: 50S ribosomal protein L28
+分子 #28: 50S ribosomal protein L29
+分子 #29: 50S ribosomal protein L30
+分子 #30: 50S ribosomal protein L32
+分子 #32: 30S ribosomal protein S2
+分子 #33: 30S ribosomal protein S3
+分子 #34: 30S ribosomal protein S4
+分子 #35: 30S ribosomal protein S5
+分子 #36: 30S ribosomal protein S6
+分子 #37: 30S ribosomal protein S7
+分子 #38: 30S ribosomal protein S8
+分子 #39: 30S ribosomal protein S9
+分子 #40: 30S ribosomal protein S10
+分子 #41: 30S ribosomal protein S11
+分子 #42: 30S ribosomal protein S12
+分子 #43: 30S ribosomal protein S13
+分子 #44: 30S ribosomal protein S14
+分子 #45: 30S ribosomal protein S15
+分子 #46: 30S ribosomal protein S16
+分子 #47: 30S ribosomal protein S17
+分子 #48: 30S ribosomal protein S18
+分子 #49: 30S ribosomal protein S19
+分子 #50: 30S ribosomal protein S20
+分子 #51: 30S ribosomal protein S21
+分子 #5: 23s ribosomal RNA
+分子 #6: 5s ribosomal RNA
+分子 #31: 16s Ribosomal RNA
+分子 #52: tRNA-met
+分子 #53: mRNA
+分子 #54: ZINC ION
+分子 #55: Eravacycline
+分子 #56: MAGNESIUM ION
+分子 #57: 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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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
緩衝液 | pH: 7.6 |
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凍結 | 凍結剤: ETHANE |
-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: 46.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: SPOT SCAN / 撮影モード: BRIGHT FIELD |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
-画像解析
初期モデル | モデルのタイプ: INSILICO MODEL / In silico モデル: ab initio |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 2.66 Å / 解像度の算出法: FSC 0.143 CUT-OFF 詳細: 50S resolution = 2.66. 30S core resolution = 2.78. 30S head resolution = 2.75. 使用した粒子像数: 40679 |
初期 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD / ソフトウェア - 名称: cryoSPARC |