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データを開く
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基本情報
| 登録情報 | ![]() | |||||||||
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| タイトル | Structure of WT E.coli ribosome 70S subunit with complexed with mRNA, P-site fMet-NH-tRNAfMet and A-site (R) beta-2-hydroxy-BocLysine acid charged NH-tRNAPyl | |||||||||
マップデータ | composite 70S map | |||||||||
試料 |
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キーワード | ribosome / non-natural monomers / beta-2-hydroxy acids / unnatural monomers | |||||||||
| 機能・相同性 | 機能・相同性情報negative regulation of cytoplasmic translational initiation / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / negative regulation of cytoplasmic translation / DnaA-L2 complex / translation repressor activity / negative regulation of translational initiation / negative regulation of DNA-templated DNA replication initiation / mRNA regulatory element binding translation repressor activity ...negative regulation of cytoplasmic translational initiation / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / negative regulation of cytoplasmic translation / DnaA-L2 complex / translation repressor activity / negative regulation of translational initiation / negative regulation of DNA-templated DNA replication initiation / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / transcription antitermination / translational initiation / response to radiation / DNA-templated transcription termination / maintenance of translational fidelity / mRNA 5'-UTR binding / large ribosomal subunit / ribosomal small subunit assembly / transferase activity / ribosome binding / ribosomal small subunit biogenesis / 5S rRNA binding / small ribosomal subunit / ribosomal large subunit assembly / small ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / negative regulation of translation / rRNA binding / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / DNA binding / RNA binding / zinc ion binding / membrane / cytoplasm / cytosol 類似検索 - 分子機能 | |||||||||
| 生物種 | ![]() Methanomethylophilus alvi (古細菌) | |||||||||
| 手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 2.14 Å | |||||||||
データ登録者 | Majumdar C / Cate JHD | |||||||||
| 資金援助 | 米国, 1件
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引用 | ジャーナル: J Am Chem Soc / 年: 2026タイトル: Co-Translational Incorporation of - and -β-Hydroxy Acids : A Structural and Biochemical Study on the Ribosome. 著者: Chandrima Majumdar / Alexandra D Kent / Noah X Hamlish / Cathy Zhu / Katelyn A Fitzgerald / Jamie H D Cate / Alanna Schepartz / ![]() 要旨: Engineering the translation apparatus to accept backbone-modified amino acid analogues would enable the programmed synthesis of sequence-defined biopolymers with tunable properties. β-Hydroxy acids ...Engineering the translation apparatus to accept backbone-modified amino acid analogues would enable the programmed synthesis of sequence-defined biopolymers with tunable properties. β-Hydroxy acids are of particular interest because they could support the programmed biosynthesis of both biocompatible polyester materials as well as natural product-like depsipeptides. Previous work has reported that both enantiomers of β-hydroxy-N-Boc-lysine (β-OH-BocK) are substrates for the orthogonal pyrrolysyl-tRNA synthetase (PylRS)/tRNA pair, but only one enantiomer is introduced into protein . Here we make use of high-resolution cryogenic electron microscopy (cryo-EM) to determine the structural basis for this observation. These structures reveal both β-OH-BocK isomers equally well-positioned within the ribosomal A site regardless of stereochemistry. Consistent with this observation, translation reactions charged with tRNAs acylated with - or -β-OH-BocK produced roughly equal amounts of translated product when quantified on the basis of either mass spectrometry or luminescence. Together, these experiments imply that the substantial preferential incorporation of one enantiomer over the other observed previously results primarily from deficiencies in the steps that precede bond formation by the ribosome. Indeed, as predicted by this work and demonstrated in an accompanying paper (Soni, C. Co-Translational Incorporation of ()- and ()-β-Hydroxyacids : Directed Evolution of Efficient Aminoacyl-tRNA Synthetases. 2026, 148, 10.1021/jacs.5c18595), when cells are provided with an active and orthogonal aminoacyl-tRNA synthetase/tRNA pair that accepts both - and -β-OH-BocK as substrates, both monomers are introduced into protein in good yield and with high fidelity. | |||||||||
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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 | emd_70051.map.gz | 63.6 MB | EMDBマップデータ形式 | |
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| ヘッダ (付随情報) | emd-70051-v30.xml emd-70051.xml | 73.7 KB 73.7 KB | 表示 表示 | EMDBヘッダ |
| FSC (解像度算出) | emd_70051_fsc.xml | 18 KB | 表示 | FSCデータファイル |
| 画像 | emd_70051.png | 85.2 KB | ||
| Filedesc metadata | emd-70051.cif.gz | 15.6 KB | ||
| アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-70051 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-70051 | HTTPS FTP |
-関連構造データ
| 関連構造データ | ![]() 9o2yMC ![]() 9o2xC C: 同じ文献を引用 ( M: このマップから作成された原子モデル |
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| 類似構造データ | 類似検索 - 機能・相同性 F&H 検索 |
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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 |
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マップ
| ファイル | ダウンロード / ファイル: emd_70051.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 | composite 70S map | ||||||||||||||||||||||||||||||||||||
| 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: 0.8235 Å | ||||||||||||||||||||||||||||||||||||
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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 : 70S ribosome particle
+超分子 #1: 70S ribosome particle
+分子 #1: 16S rRNA
+分子 #22: mRNA
+分子 #23: P-site tRNA fMet
+分子 #24: 23S rRNA
+分子 #25: 5S rRNA
+分子 #55: A-site tRNAPyl
+分子 #2: 30S ribosomal protein S2
+分子 #3: Small ribosomal subunit protein uS3
+分子 #4: Small ribosomal subunit protein uS4
+分子 #5: Small ribosomal subunit protein uS5
+分子 #6: Small ribosomal subunit protein bS6
+分子 #7: Small ribosomal subunit protein uS7
+分子 #8: Small ribosomal subunit protein uS8
+分子 #9: Small ribosomal subunit protein uS9
+分子 #10: Small ribosomal subunit protein uS10
+分子 #11: Small ribosomal subunit protein uS11
+分子 #12: Small ribosomal subunit protein uS12
+分子 #13: Small ribosomal subunit protein uS13
+分子 #14: Small ribosomal subunit protein uS14
+分子 #15: Small ribosomal subunit protein uS15
+分子 #16: Small ribosomal subunit protein bS16
+分子 #17: Small ribosomal subunit protein uS17
+分子 #18: Small ribosomal subunit protein bS18
+分子 #19: Small ribosomal subunit protein uS19
+分子 #20: Small ribosomal subunit protein bS20
+分子 #21: Small ribosomal subunit protein bS21
+分子 #26: 50S ribosomal protein L2
+分子 #27: 50S ribosomal protein L3
+分子 #28: Large ribosomal subunit protein uL4
+分子 #29: Large ribosomal subunit protein uL5
+分子 #30: Large ribosomal subunit protein uL6
+分子 #31: Large ribosomal subunit protein bL9
+分子 #32: Large ribosomal subunit protein uL13
+分子 #33: Large ribosomal subunit protein uL14
+分子 #34: Large ribosomal subunit protein uL15
+分子 #35: 50S ribosomal protein L16
+分子 #36: Large ribosomal subunit protein bL17
+分子 #37: Large ribosomal subunit protein uL18
+分子 #38: Large ribosomal subunit protein bL19
+分子 #39: 50S ribosomal protein L20
+分子 #40: Large ribosomal subunit protein bL21
+分子 #41: Large ribosomal subunit protein uL22
+分子 #42: 50S ribosomal protein L23
+分子 #43: 50S ribosomal protein L24
+分子 #44: Large ribosomal subunit protein bL25
+分子 #45: Large ribosomal subunit protein bL27
+分子 #46: 50S ribosomal protein L28
+分子 #47: Large ribosomal subunit protein uL29
+分子 #48: 50S ribosomal protein L30
+分子 #49: 50S ribosomal protein L32
+分子 #50: 50S ribosomal protein L33
+分子 #51: 50S ribosomal protein L34
+分子 #52: 50S ribosomal protein L35
+分子 #53: 50S ribosomal protein L36
+分子 #54: 50S ribosomal protein L31
+分子 #56: PAROMOMYCIN
+分子 #57: MAGNESIUM ION
+分子 #58: SPERMIDINE
+分子 #59: N-FORMYLMETHIONINE
+分子 #60: SPERMINE
+分子 #61: POTASSIUM ION
+分子 #62: ZINC ION
+分子 #63: (2R)-6-[(tert-butoxycarbonyl)amino]-2-(hydroxymethyl)hexanoic acid
+分子 #64: 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 |
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試料調製
| 緩衝液 | pH: 7.5 |
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| グリッド | モデル: Quantifoil R1.2/1.3 / 材質: GOLD / メッシュ: 300 / 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: 2 / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 12 sec. |
| 凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 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 BIOQUANTUM (6k x 4k) 平均電子線量: 40.0 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: OTHER / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 1.5 µm / 最小 デフォーカス(公称値): 0.5 µm |
| 実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
ムービー
コントローラー
万見について




キーワード
Methanomethylophilus alvi (古細菌)
データ登録者
米国, 1件
引用












Z (Sec.)
Y (Row.)
X (Col.)

























解析
FIELD EMISSION GUN


