+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-5117 | |||||||||
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タイトル | Structure of furin-cleaved immature dengue virus at low pH | |||||||||
マップデータ | Immature dengue virus after furin cleavage at low pH | |||||||||
試料 |
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キーワード | dengue / prM / E / furin / maturation | |||||||||
機能・相同性 | 機能・相同性情報 flavivirin / symbiont-mediated suppression of host JAK-STAT cascade via inhibition of host TYK2 activity / host cell mitochondrion / symbiont-mediated suppression of host JAK-STAT cascade via inhibition of STAT2 activity / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of MAVS activity / double-stranded RNA binding / nucleoside-triphosphate phosphatase / channel activity / viral capsid / monoatomic ion transmembrane transport ...flavivirin / symbiont-mediated suppression of host JAK-STAT cascade via inhibition of host TYK2 activity / host cell mitochondrion / symbiont-mediated suppression of host JAK-STAT cascade via inhibition of STAT2 activity / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of MAVS activity / double-stranded RNA binding / nucleoside-triphosphate phosphatase / channel activity / viral capsid / monoatomic ion transmembrane transport / mRNA (guanine-N7)-methyltransferase / viral nucleocapsid / methyltransferase cap1 / clathrin-dependent endocytosis of virus by host cell / mRNA (nucleoside-2'-O-)-methyltransferase activity / mRNA 5'-cap (guanine-N7-)-methyltransferase activity / RNA helicase activity / host cell perinuclear region of cytoplasm / protein dimerization activity / host cell endoplasmic reticulum membrane / RNA helicase / symbiont-mediated suppression of host type I interferon-mediated signaling pathway / symbiont-mediated suppression of host innate immune response / induction by virus of host autophagy / RNA-directed RNA polymerase / viral RNA genome replication / serine-type endopeptidase activity / RNA-dependent RNA polymerase activity / virus-mediated perturbation of host defense response / fusion of virus membrane with host endosome membrane / viral envelope / host cell nucleus / virion attachment to host cell / structural molecule activity / virion membrane / ATP hydrolysis activity / proteolysis / extracellular region / ATP binding / membrane / metal ion binding 類似検索 - 分子機能 | |||||||||
生物種 | Dengue virus (デング熱ウイルス) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 22.0 Å | |||||||||
データ登録者 | Yu I / Holdaway HA / Chipman PR / Kuhn RJ / Rossmann MG / Chen J | |||||||||
引用 | ジャーナル: J Virol / 年: 2009 タイトル: Association of the pr peptides with dengue virus at acidic pH blocks membrane fusion. 著者: I-M Yu / H A Holdaway / P R Chipman / R J Kuhn / M G Rossmann / J Chen / 要旨: Flavivirus assembles into an inert particle that requires proteolytic activation by furin to enable transmission to other hosts. We previously showed that immature virus undergoes a conformational ...Flavivirus assembles into an inert particle that requires proteolytic activation by furin to enable transmission to other hosts. We previously showed that immature virus undergoes a conformational change at low pH that renders it accessible to furin (I. M. Yu, W. Zhang, H. A. Holdaway, L. Li, V. A. Kostyuchenko, P. R. Chipman, R. J. Kuhn, M. G. Rossmann, and J. Chen, Science 319:1834-1837, 2008). Here we show, using cryoelectron microscopy, that the structure of immature dengue virus at pH 6.0 is essentially the same before and after the cleavage of prM. The structure shows that after cleavage, the proteolytic product pr remains associated with the virion at acidic pH, and that furin cleavage by itself does not induce any major conformational changes. We also show by liposome cofloatation experiments that pr retention prevents membrane insertion, suggesting that pr is present on the virion in the trans-Golgi network to protect the progeny virus from fusion within the host cell. | |||||||||
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_5117.map.gz | 3.9 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-5117-v30.xml emd-5117.xml | 9.1 KB 9.1 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_5117_1.png | 176.4 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-5117 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-5117 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_5117_validation.pdf.gz | 319.5 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_5117_full_validation.pdf.gz | 319 KB | 表示 | |
XML形式データ | emd_5117_validation.xml.gz | 5.8 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-5117 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-5117 | 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_5117.map.gz / 形式: CCP4 / 大きさ: 9.2 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Immature dengue virus after furin cleavage at low pH | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 4.4 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
-試料の構成要素
-全体 : Immature dengue virus at low pH after furin cleavage
全体 | 名称: Immature dengue virus at low pH after furin cleavage |
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要素 |
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-超分子 #1000: Immature dengue virus at low pH after furin cleavage
超分子 | 名称: Immature dengue virus at low pH after furin cleavage タイプ: sample / ID: 1000 / Number unique components: 2 |
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-超分子 #1: Dengue virus
超分子 | 名称: Dengue virus / タイプ: virus / ID: 1 / Name.synonym: dengue / NCBI-ID: 12637 / 生物種: Dengue virus / データベース: NCBI / ウイルスタイプ: VIRION / ウイルス・単離状態: STRAIN / ウイルス・エンベロープ: Yes / ウイルス・中空状態: No / Syn species name: dengue |
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宿主 | 別称: VERTEBRATES |
ウイルス殻 | Shell ID: 1 / T番号(三角分割数): 3 |
-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: 6 / 詳細: 6 mM Tris-HCL, 25 mM MES, 120 mM NaCl, 1 mM EDTA |
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凍結 | 凍結剤: ETHANE / 装置: OTHER 手法: A small vial of ethane is placed inside a larger liquid nitrogen reservoir. The grid holding a few microliters of the sample is held in place at the bottom of a plunger by the means of fine ...手法: A small vial of ethane is placed inside a larger liquid nitrogen reservoir. The grid holding a few microliters of the sample is held in place at the bottom of a plunger by the means of fine tweezers. Once the ethane in the vial is completely frozen, it needs to be slightly melted. When the liquid ethane is ready, a piece of filter paper is then pressed against the sample to blot of excess buffer, sufficient to leave a thin layer on the grid. After a predetermined time, the filter paper is removed, and the plunger is allowed to drop into the liquid ethane. Once the grid enters the liquid ethane, the sample is rapidly frozen, and the grid is transferred under liquid nitrogen to a storage box immersed liquid nitrogen for later use in the microscope. |
-電子顕微鏡法
顕微鏡 | FEI/PHILIPS CM200FEG |
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日付 | 2008年6月13日 |
撮影 | カテゴリ: CCD / フィルム・検出器のモデル: GATAN MULTISCAN / 実像数: 45 / 平均電子線量: 25 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 3.8 µm / 最小 デフォーカス(公称値): 1.8 µm / 倍率(公称値): 50000 |
試料ステージ | 試料ホルダー: Eucentric / 試料ホルダーモデル: GATAN LIQUID NITROGEN |
-画像解析
CTF補正 | 詳細: Each particle |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 22.0 Å / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア - 名称: EM3DR / 使用した粒子像数: 635 |