+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 7n7u | ||||||
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タイトル | Crystal Structure of SARS-CoV-2 NendoU in complex with LIZA-7 | ||||||
要素 | Uridylate-specific endoribonuclease | ||||||
キーワード | VIRAL PROTEIN / SGC - Diamond I04-1 fragment screening / XChemExplorer / nsp15 / nendoU / sars-cov-2 / sars / covid / covid19 | ||||||
機能・相同性 | 機能・相同性情報 protein guanylyltransferase activity / RNA endonuclease activity, producing 3'-phosphomonoesters / mRNA guanylyltransferase activity / 5'-3' RNA helicase activity / 付加脱離酵素(リアーゼ); P-Oリアーゼ類; - / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of TBK1 activity / Assembly of the SARS-CoV-2 Replication-Transcription Complex (RTC) / Maturation of replicase proteins / ISG15-specific peptidase activity / Transcription of SARS-CoV-2 sgRNAs ...protein guanylyltransferase activity / RNA endonuclease activity, producing 3'-phosphomonoesters / mRNA guanylyltransferase activity / 5'-3' RNA helicase activity / 付加脱離酵素(リアーゼ); P-Oリアーゼ類; - / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of TBK1 activity / Assembly of the SARS-CoV-2 Replication-Transcription Complex (RTC) / Maturation of replicase proteins / ISG15-specific peptidase activity / Transcription of SARS-CoV-2 sgRNAs / Translation of Replicase and Assembly of the Replication Transcription Complex / TRAF3-dependent IRF activation pathway / Replication of the SARS-CoV-2 genome / snRNP Assembly / double membrane vesicle viral factory outer membrane / 加水分解酵素; エステル加水分解酵素; 5'-リン酸モノエステル産生エキソリボヌクレアーゼ / 5'-3' DNA helicase activity / SARS coronavirus main proteinase / host cell endosome / host cell endoplasmic reticulum-Golgi intermediate compartment / 3'-5'-RNA exonuclease activity / symbiont-mediated suppression of host toll-like receptor signaling pathway / symbiont-mediated degradation of host mRNA / mRNA guanylyltransferase / symbiont-mediated suppression of host ISG15-protein conjugation / G-quadruplex RNA binding / SARS-CoV-2 modulates host translation machinery / omega peptidase activity / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of IRF3 activity / mRNA (guanine-N7)-methyltransferase / host cell Golgi apparatus / methyltransferase cap1 / symbiont-mediated perturbation of host ubiquitin-like protein modification / mRNA (nucleoside-2'-O-)-methyltransferase activity / DNA helicase / mRNA 5'-cap (guanine-N7-)-methyltransferase activity / ubiquitinyl hydrolase 1 / cysteine-type deubiquitinase activity / 加水分解酵素; プロテアーゼ; ペプチド結合加水分解酵素; システインプロテアーゼ / single-stranded RNA binding / host cell endoplasmic reticulum membrane / host cell perinuclear region of cytoplasm / viral protein processing / lyase activity / symbiont-mediated suppression of host type I interferon-mediated signaling pathway / RNA helicase / induction by virus of host autophagy / copper ion binding / RNA-directed RNA polymerase / viral RNA genome replication / cysteine-type endopeptidase activity / RNA-dependent RNA polymerase activity / virus-mediated perturbation of host defense response / DNA-templated transcription / lipid binding / host cell nucleus / SARS-CoV-2 activates/modulates innate and adaptive immune responses / ATP hydrolysis activity / proteolysis / RNA binding / zinc ion binding / ATP binding / membrane 類似検索 - 分子機能 | ||||||
生物種 | Severe acute respiratory syndrome coronavirus 2 (ウイルス) | ||||||
手法 | X線回折 / シンクロトロン / 分子置換 / 解像度: 2.06 Å | ||||||
データ登録者 | Godoy, A.S. / Douangamath, A. / Nakamura, A.M. / Dias, A. / Krojer, T. / Noske, G.D. / Gawiljuk, V.O. / Fernandes, R.S. / Fairhead, M. / Powell, A. ...Godoy, A.S. / Douangamath, A. / Nakamura, A.M. / Dias, A. / Krojer, T. / Noske, G.D. / Gawiljuk, V.O. / Fernandes, R.S. / Fairhead, M. / Powell, A. / Dunnet, L. / Aimon, A. / Fearon, D. / Brandao-Neto, J. / Skyner, R. / von Delft, F. / Oliva, G. | ||||||
資金援助 | ブラジル, 1件
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引用 | ジャーナル: Nucleic Acids Res / 年: 2023 タイトル: Allosteric regulation and crystallographic fragment screening of SARS-CoV-2 NSP15 endoribonuclease. 著者: Andre Schutzer Godoy / Aline Minalli Nakamura / Alice Douangamath / Yun Song / Gabriela Dias Noske / Victor Oliveira Gawriljuk / Rafaela Sachetto Fernandes / Humberto D Muniz Pereira / ...著者: Andre Schutzer Godoy / Aline Minalli Nakamura / Alice Douangamath / Yun Song / Gabriela Dias Noske / Victor Oliveira Gawriljuk / Rafaela Sachetto Fernandes / Humberto D Muniz Pereira / Ketllyn Irene Zagato Oliveira / Daren Fearon / Alexandre Dias / Tobias Krojer / Michael Fairhead / Alisa Powell / Louise Dunnet / Jose Brandao-Neto / Rachael Skyner / Rod Chalk / Dávid Bajusz / Miklós Bege / Anikó Borbás / György Miklós Keserű / Frank von Delft / Glaucius Oliva / 要旨: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19). The NSP15 endoribonuclease enzyme, known as NendoU, is highly conserved and ...Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19). The NSP15 endoribonuclease enzyme, known as NendoU, is highly conserved and plays a critical role in the ability of the virus to evade the immune system. NendoU is a promising target for the development of new antiviral drugs. However, the complexity of the enzyme's structure and kinetics, along with the broad range of recognition sequences and lack of structural complexes, hampers the development of inhibitors. Here, we performed enzymatic characterization of NendoU in its monomeric and hexameric form, showing that hexamers are allosteric enzymes with a positive cooperative index, and with no influence of manganese on enzymatic activity. Through combining cryo-electron microscopy at different pHs, X-ray crystallography and biochemical and structural analysis, we showed that NendoU can shift between open and closed forms, which probably correspond to active and inactive states, respectively. We also explored the possibility of NendoU assembling into larger supramolecular structures and proposed a mechanism for allosteric regulation. In addition, we conducted a large fragment screening campaign against NendoU and identified several new allosteric sites that could be targeted for the development of new inhibitors. Overall, our findings provide insights into the complex structure and function of NendoU and offer new opportunities for the development of inhibitors. | ||||||
履歴 |
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-構造の表示
構造ビューア | 分子: MolmilJmol/JSmol |
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-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 7n7u.cif.gz | 163.9 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb7n7u.ent.gz | 127.7 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 7n7u.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
その他 | その他のダウンロード |
-検証レポート
文書・要旨 | 7n7u_validation.pdf.gz | 1.1 MB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 7n7u_full_validation.pdf.gz | 1.1 MB | 表示 | |
XML形式データ | 7n7u_validation.xml.gz | 32.9 KB | 表示 | |
CIF形式データ | 7n7u_validation.cif.gz | 48.2 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/n7/7n7u ftp://data.pdbj.org/pub/pdb/validation_reports/n7/7n7u | HTTPS FTP |
-関連構造データ
関連構造データ | 5s6xC 5s6yC 5s6zC 5s70C 5s71C 5s72C 5sa4C 5sa5C 5sa6C 5sa7C 5sa8C 5sa9C 5saaC 5sabC 5sacC 5sadC 5saeC 5safC 5sagC 5sahC 5saiC 5sbfC 7kegC 7kehC 7kf4C 7me0C 7n7rC 7n7wC 7n7yC 7n83C 7rb0C 7rb2C 6x1bS S: 精密化の開始モデル C: 同じ文献を引用 (文献) |
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類似構造データ |
-リンク
-集合体
登録構造単位 |
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単位格子 |
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-要素
#1: タンパク質 | 分子量: 39112.535 Da / 分子数: 2 / 由来タイプ: 組換発現 由来: (組換発現) Severe acute respiratory syndrome coronavirus 2 (ウイルス) 遺伝子: rep, 1a-1b / 発現宿主: Escherichia coli (大腸菌) 参照: UniProt: P0DTD1, 加水分解酵素; エステル加水分解酵素 #2: 化合物 | #3: 水 | ChemComp-HOH / | 研究の焦点であるリガンドがあるか | Y | |
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-実験情報
-実験
実験 | 手法: X線回折 / 使用した結晶の数: 1 |
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-試料調製
結晶 | マシュー密度: 4.56 Å3/Da / 溶媒含有率: 73 % |
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結晶化 | 温度: 293 K / 手法: 蒸気拡散法 / 詳細: '0.1 M Na3 Citrate pH 5,14% w/v PEG6000 |
-データ収集
回折 | 平均測定温度: 100 K / Serial crystal experiment: N | ||||||||||||||||||||||||||||||
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放射光源 | 由来: シンクロトロン / サイト: Diamond / ビームライン: I04-1 / 波長: 0.91256 Å | ||||||||||||||||||||||||||||||
検出器 | タイプ: DECTRIS PILATUS 6M / 検出器: PIXEL / 日付: 2020年7月7日 | ||||||||||||||||||||||||||||||
放射 | プロトコル: SINGLE WAVELENGTH / 単色(M)・ラウエ(L): M / 散乱光タイプ: x-ray | ||||||||||||||||||||||||||||||
放射波長 | 波長: 0.91256 Å / 相対比: 1 | ||||||||||||||||||||||||||||||
反射 | 解像度: 1.96→129.642 Å / Num. obs: 100329 / % possible obs: 94.3 % / 冗長度: 10.2 % / Biso Wilson estimate: 35.87 Å2 / CC1/2: 0.994 / Rmerge(I) obs: 0.188 / Rpim(I) all: 0.062 / Rrim(I) all: 0.198 / Net I/σ(I): 8.5 | ||||||||||||||||||||||||||||||
反射 シェル | Diffraction-ID: 1
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-解析
ソフトウェア |
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精密化 | 構造決定の手法: 分子置換 開始モデル: 6x1b 解像度: 2.06→64.97 Å / SU ML: 0.36 / 交差検証法: THROUGHOUT / σ(F): 1.34 / 位相誤差: 30 / 立体化学のターゲット値: ML
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溶媒の処理 | 減衰半径: 0.9 Å / VDWプローブ半径: 1.11 Å / 溶媒モデル: FLAT BULK SOLVENT MODEL | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
原子変位パラメータ | Biso max: 105.07 Å2 / Biso mean: 40.8109 Å2 / Biso min: 23.16 Å2 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
精密化ステップ | サイクル: final / 解像度: 2.06→64.97 Å
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LS精密化 シェル | Refine-ID: X-RAY DIFFRACTION / Rfactor Rfree error: 0 / Total num. of bins used: 27
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