+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-10656 | ||||||||||||||||||
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タイトル | E. coli 70S ribosome in complex with dirithromycin, and deacylated tRNA(iMet) (focused classification). | ||||||||||||||||||
マップデータ | 70S ribosome in complex with dirithromycin and deacylated tRNAiMet in the P site | ||||||||||||||||||
試料 |
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キーワード | 70S ribosome / dirithromycin / antibiotics / cryo-EM / RIBOSOME | ||||||||||||||||||
機能・相同性 | 機能・相同性情報 stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis ...stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / negative regulation of cytoplasmic translation / translational termination / four-way junction DNA binding / DnaA-L2 complex / translation repressor activity / negative regulation of DNA-templated DNA replication initiation / negative regulation of translational initiation / regulation of mRNA stability / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / transcription elongation factor complex / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / DNA endonuclease activity / response to reactive oxygen species / transcription antitermination / regulation of cell growth / DNA-templated transcription termination / maintenance of translational fidelity / response to radiation / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosome biogenesis / ribosome binding / regulation of translation / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / molecular adaptor activity / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / DNA binding / RNA binding / zinc ion binding / membrane / cytosol / cytoplasm 類似検索 - 分子機能 | ||||||||||||||||||
生物種 | Escherichia coli (strain K12) (大腸菌) / Escherichia coli K-12 (大腸菌) | ||||||||||||||||||
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 | Pichkur EB / Polikanov YS | ||||||||||||||||||
資金援助 | ロシア, 米国, 5件
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引用 | ジャーナル: RNA / 年: 2020 タイトル: Insights into the improved macrolide inhibitory activity from the high-resolution cryo-EM structure of dirithromycin bound to the 70S ribosome. 著者: Evgeny B Pichkur / Alena Paleskava / Andrey G Tereshchenkov / Pavel Kasatsky / Ekaterina S Komarova / Dmitrii I Shiriaev / Alexey A Bogdanov / Olga A Dontsova / Ilya A Osterman / Petr V ...著者: Evgeny B Pichkur / Alena Paleskava / Andrey G Tereshchenkov / Pavel Kasatsky / Ekaterina S Komarova / Dmitrii I Shiriaev / Alexey A Bogdanov / Olga A Dontsova / Ilya A Osterman / Petr V Sergiev / Yury S Polikanov / Alexander G Myasnikov / Andrey L Konevega / 要旨: Macrolides are one of the most successful and widely used classes of antibacterials, which kill or stop the growth of pathogenic bacteria by binding near the active site of the ribosome and ...Macrolides are one of the most successful and widely used classes of antibacterials, which kill or stop the growth of pathogenic bacteria by binding near the active site of the ribosome and interfering with protein synthesis. Dirithromycin is a derivative of the prototype macrolide erythromycin with additional hydrophobic side chain. In our recent study, we have discovered that the side chain of dirithromycin forms lone pair-π stacking interaction with the aromatic imidazole ring of the His69 residue in ribosomal protein uL4 of the 70S ribosome. In the current work, we found that neither the presence of the side chain, nor the additional contact with the ribosome, improve the binding affinity of dirithromycin to the ribosome. Nevertheless, we found that dirithromycin is a more potent inhibitor of in vitro protein synthesis in comparison with its parent compound, erythromycin. Using high-resolution cryo-electron microscopy, we determined the structure of the dirithromycin bound to the translating 70S ribosome, which suggests that the better inhibitory properties of the drug could be rationalized by the side chain of dirithromycin pointing into the lumen of the nascent peptide exit tunnel, where it can interfere with the normal passage of the growing polypeptide chain. | ||||||||||||||||||
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_10656.map.gz | 467.3 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-10656-v30.xml emd-10656.xml | 70.3 KB 70.3 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_10656.png | 285.1 KB | ||
Filedesc metadata | emd-10656.cif.gz | 14.4 KB | ||
その他 | emd_10656_additional_1.map.gz | 466.7 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-10656 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-10656 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_10656_validation.pdf.gz | 190.5 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_10656_full_validation.pdf.gz | 190 KB | 表示 | |
XML形式データ | emd_10656_validation.xml.gz | 500 B | 表示 | |
CIF形式データ | emd_10656_validation.cif.gz | 374 B | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10656 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-10656 | 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_10656.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 | 70S ribosome in complex with dirithromycin and deacylated tRNAiMet in the P site | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 0.86 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: 70S ribosome in complex with dirithromycin and deacylated...
ファイル | emd_10656_additional_1.map | ||||||||||||
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注釈 | 70S ribosome in complex with dirithromycin and deacylated tRNAiMet in the P site. Sharpened map. | ||||||||||||
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 : E. coli 70S ribosome in complex with dirithromycin and deacylated...
+超分子 #1: E. coli 70S ribosome in complex with dirithromycin and deacylated...
+分子 #1: 16S rRNA
+分子 #22: 23S rRNA
+分子 #23: 5S rRNA
+分子 #53: Deacylated tRNAi(Met)
+分子 #2: 30S ribosomal protein S2
+分子 #3: 30S ribosomal protein S3
+分子 #4: 30S ribosomal protein S4
+分子 #5: 30S ribosomal protein S5
+分子 #6: 30S ribosomal protein S6
+分子 #7: 30S ribosomal protein S7
+分子 #8: 30S ribosomal protein S8
+分子 #9: 30S ribosomal protein S9
+分子 #10: 30S ribosomal protein S10
+分子 #11: 30S ribosomal protein S11
+分子 #12: 30S ribosomal protein S12
+分子 #13: 30S ribosomal protein S13
+分子 #14: 30S ribosomal protein S14
+分子 #15: 30S ribosomal protein S15
+分子 #16: 30S ribosomal protein S16
+分子 #17: 30S ribosomal protein S17
+分子 #18: 30S ribosomal protein S18
+分子 #19: 30S ribosomal protein S19
+分子 #20: 30S ribosomal protein S20
+分子 #21: 30S ribosomal protein S21
+分子 #24: 50S ribosomal protein L2
+分子 #25: 50S ribosomal protein L3
+分子 #26: 50S ribosomal protein L4
+分子 #27: 50S ribosomal protein L5
+分子 #28: 50S ribosomal protein L6
+分子 #29: 50S ribosomal protein L10
+分子 #30: 50S ribosomal protein L11
+分子 #31: 50S ribosomal protein L13
+分子 #32: 50S ribosomal protein L14
+分子 #33: 50S ribosomal protein L15
+分子 #34: 50S ribosomal protein L16
+分子 #35: 50S ribosomal protein L17
+分子 #36: 50S ribosomal protein L18
+分子 #37: 50S ribosomal protein L19
+分子 #38: 50S ribosomal protein L20
+分子 #39: 50S ribosomal protein L21
+分子 #40: 50S ribosomal protein L22
+分子 #41: 50S ribosomal protein L23
+分子 #42: 50S ribosomal protein L24
+分子 #43: 50S ribosomal protein L25
+分子 #44: 50S ribosomal protein L27
+分子 #45: 50S ribosomal protein L28
+分子 #46: 50S ribosomal protein L29
+分子 #47: 50S ribosomal protein L30
+分子 #48: 50S ribosomal protein L32
+分子 #49: 50S ribosomal protein L33
+分子 #50: 50S ribosomal protein L34
+分子 #51: 50S ribosomal protein L35
+分子 #52: 50S ribosomal protein L36
+分子 #54: Dirithromycin
-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: 7.5 |
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グリッド | モデル: Quantifoil R2/2 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: CONTINUOUS / 支持フィルム - Film thickness: 10 / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 45 sec. / 前処理 - 雰囲気: AIR |
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: FEI FALCON II (4k x 4k) 検出モード: INTEGRATING / デジタル化 - 画像ごとのフレーム数: 2-27 / 平均露光時間: 1.4 sec. / 平均電子線量: 80.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 100.0 µm / 最大 デフォーカス(補正後): 2.2 µm / 最小 デフォーカス(補正後): 0.3 µm / 照射モード: SPOT SCAN / 撮影モード: BRIGHT FIELD / Cs: 0.1 mm / 最大 デフォーカス(公称値): 2.2 µm / 最小 デフォーカス(公称値): 0.3 µm / 倍率(公称値): 75000 |
試料ステージ | ホルダー冷却材: NITROGEN |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |