- EMDB-5141: Ab initio reconstruction of the E. coli 70S ribosome complex (70S... -
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基本情報
登録情報
データベース: EMDB / ID: EMD-5141
タイトル
Ab initio reconstruction of the E. coli 70S ribosome complex (70S-fMet-tRNAfMet-Phe-tRNAPhe-EF-Tu-GDP-kirromycin) via the asymmetric random-model method.
マップデータ
This is a ab initio starting model for the E. coli 70S ribosome complex (70S-fMet-tRNAfMet-Phe-tRNAPhe-EF-Tu-GDP-kirromycin)
試料
試料: E. coli 70S ribosome complex, 70S-fMet-tRNAfMet-Phe-tRNAPhe-EF-Tu-GDP-kirromycin
複合体: 70S ribosome complex
キーワード
Random-model method / ab initio reconstruction / E. coli 70S ribosome
ジャーナル: Nat Struct Biol / 年: 2003 タイトル: Incorporation of aminoacyl-tRNA into the ribosome as seen by cryo-electron microscopy. 著者: Mikel Valle / Andrey Zavialov / Wen Li / Scott M Stagg / Jayati Sengupta / Rikke C Nielsen / Poul Nissen / Stephen C Harvey / Måns Ehrenberg / Joachim Frank / 要旨: Aminoacyl-tRNAs (aa-tRNAs) are delivered to the ribosome as part of the ternary complex of aa-tRNA, elongation factor Tu (EF-Tu) and GTP. Here, we present a cryo-electron microscopy (cryo-EM) study, ...Aminoacyl-tRNAs (aa-tRNAs) are delivered to the ribosome as part of the ternary complex of aa-tRNA, elongation factor Tu (EF-Tu) and GTP. Here, we present a cryo-electron microscopy (cryo-EM) study, at a resolution of approximately 9 A, showing that during the incorporation of the aa-tRNA into the 70S ribosome of Escherichia coli, the flexibility of aa-tRNA allows the initial codon recognition and its accommodation into the ribosomal A site. In addition, a conformational change observed in the GTPase-associated center (GAC) of the ribosomal 50S subunit may provide the mechanism by which the ribosome promotes a relative movement of the aa-tRNA with respect to EF-Tu. This relative rearrangement seems to facilitate codon recognition by the incoming aa-tRNA, and to provide the codon-anticodon recognition-dependent signal for the GTPase activity of EF-Tu. From these new findings we propose a mechanism that can explain the sequence of events during the decoding of mRNA on the ribosome.
全体 : E. coli 70S ribosome complex, 70S-fMet-tRNAfMet-Phe-tRNAPhe-EF-Tu...
全体
名称: E. coli 70S ribosome complex, 70S-fMet-tRNAfMet-Phe-tRNAPhe-EF-Tu-GDP-kirromycin
要素
試料: E. coli 70S ribosome complex, 70S-fMet-tRNAfMet-Phe-tRNAPhe-EF-Tu-GDP-kirromycin
複合体: 70S ribosome complex
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超分子 #1000: E. coli 70S ribosome complex, 70S-fMet-tRNAfMet-Phe-tRNAPhe-EF-Tu...
超分子
名称: E. coli 70S ribosome complex, 70S-fMet-tRNAfMet-Phe-tRNAPhe-EF-Tu-GDP-kirromycin タイプ: sample / ID: 1000 / 集合状態: One ribosome binds two tRNA and one EF-Tu / Number unique components: 6
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超分子 #1: 70S ribosome complex
超分子
名称: 70S ribosome complex / タイプ: complex / ID: 1 / 組換発現: No / データベース: NCBI / Ribosome-details: ribosome-prokaryote: ALL
由来(天然)
生物種: Escherichia coli (大腸菌)
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実験情報
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構造解析
手法
クライオ電子顕微鏡法
解析
単粒子再構成法
試料の集合状態
particle
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試料調製
凍結
凍結剤: ETHANE / 装置: OTHER
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電子顕微鏡法
顕微鏡
FEI TECNAI F20
撮影
カテゴリ: FILM / フィルム・検出器のモデル: GENERIC FILM / デジタル化 - スキャナー: ZEISS SCAI / デジタル化 - サンプリング間隔: 14 µm
試料ホルダー: Eucentric / 試料ホルダーモデル: SIDE ENTRY, EUCENTRIC
実験機器
モデル: Tecnai F20 / 画像提供: FEI Company
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画像解析
CTF補正
詳細: Phase-flipped per micrograph
最終 再構成
アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 23.0 Å / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア - 名称: PFT3DR, Bsoft 詳細: Random-model method. Angular step-size was initially set to 20 deg. in the first iteration and gradually decreased by 0.19 deg. in each successive iteration, until a lower limit of 1 deg. was reached. 使用した粒子像数: 10000