+
データを開く
-
基本情報
登録情報 | データベース: EMDB / ID: EMD-5997 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
タイトル | Electron cryo-microscopy of chimeric HT-VP2-466 capsid | |||||||||
![]() | Reconstruction of mutant IBDV (chimeric HT-VP2-466 capsid) | |||||||||
![]() |
| |||||||||
![]() | chimeric virus-like particles / IBDV VLP / Cryomicroscopy | |||||||||
生物種 | ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 20.8 Å | |||||||||
![]() | Pascual E / Mata CP / Gomez-Blanco J / Carrascosa JL / Caston JR | |||||||||
![]() | ![]() タイトル: Structural basis for the development of avian virus capsids that display influenza virus proteins and induce protective immunity. 著者: Elena Pascual / Carlos P Mata / Josué Gómez-Blanco / Noelia Moreno / Juan Bárcena / Esther Blanco / Ariel Rodríguez-Frandsen / Amelia Nieto / José L Carrascosa / José R Castón / ![]() 要旨: Bioengineering of viruses and virus-like particles (VLPs) is a well-established approach in the development of new and improved vaccines against viral and bacterial pathogens. We report here that the ...Bioengineering of viruses and virus-like particles (VLPs) is a well-established approach in the development of new and improved vaccines against viral and bacterial pathogens. We report here that the capsid of a major avian pathogen, infectious bursal disease virus (IBDV), can accommodate heterologous proteins to induce protective immunity. The structural units of the ~70-nm-diameter T=13 IBDV capsid are trimers of VP2, which is made as a precursor (pVP2). The pVP2 C-terminal domain has an amphipathic α helix that controls VP2 polymorphism. In the absence of the VP3 scaffolding protein, 466-residue pVP2 intermediates bearing this α helix assemble into genuine VLPs only when expressed with an N-terminal His6 tag (the HT-VP2-466 protein). HT-VP2-466 capsids are optimal for protein insertion, as they are large enough (cargo space, ~78,000 nm(3)) and are assembled from a single protein. We explored HT-VP2-466-based chimeric capsids initially using enhanced green fluorescent protein (EGFP). The VLP assembly yield was efficient when we coexpressed EGFP-HT-VP2-466 and HT-VP2-466 from two recombinant baculoviruses. The native EGFP structure (~240 copies/virion) was successfully inserted in a functional form, as VLPs were fluorescent, and three-dimensional cryo-electron microscopy showed that the EGFP molecules incorporated at the inner capsid surface. Immunization of mice with purified EGFP-VLPs elicited anti-EGFP antibodies. We also inserted hemagglutinin (HA) and matrix (M2) protein epitopes derived from the mouse-adapted A/PR/8/34 influenza virus and engineered several HA- and M2-derived chimeric capsids. Mice immunized with VLPs containing the HA stalk, an M2 fragment, or both antigens developed full protection against viral challenge. IMPORTANCE: Virus-like particles (VLPs) are multimeric protein cages that mimic the infectious virus capsid and are potential candidates as nonliving vaccines that induce long-lasting protection. ...IMPORTANCE: Virus-like particles (VLPs) are multimeric protein cages that mimic the infectious virus capsid and are potential candidates as nonliving vaccines that induce long-lasting protection. Chimeric VLPs can display or include foreign antigens, which could be a conserved epitope to elicit broadly neutralizing antibodies or several variable epitopes effective against a large number of viral strains. We report the biochemical, structural, and immunological characterization of chimeric VLPs derived from infectious bursal disease virus (IBDV), an important poultry pathogen. To test the potential of IBDV VLPs as a vaccine vehicle, we used the enhanced green fluorescent protein and two fragments derived from the hemagglutinin and the M2 matrix protein of the human murine-adapted influenza virus. The IBDV capsid protein fused to influenza virus peptides formed assemblies able to protect mice against viral challenge. Our studies establish the basis for a new generation of multivalent IBDV-based vaccines. | |||||||||
履歴 |
|
-
構造の表示
ムービー |
![]() |
---|---|
構造ビューア | EMマップ: ![]() ![]() ![]() |
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 | ![]() | 82.5 MB | ![]() | |
---|---|---|---|---|
ヘッダ (付随情報) | ![]() ![]() | 9.1 KB 9.1 KB | 表示 表示 | ![]() |
画像 | ![]() ![]() | 51.6 KB 4.5 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
-
リンク
EMDBのページ | ![]() ![]() |
---|
-
マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Reconstruction of mutant IBDV (chimeric HT-VP2-466 capsid) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 2.8 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
|
-添付データ
-
試料の構成要素
-全体 : Infectious bursal disease virus mutant HT-VP2-466
全体 | 名称: Infectious bursal disease virus mutant HT-VP2-466 |
---|---|
要素 |
|
-超分子 #1000: Infectious bursal disease virus mutant HT-VP2-466
超分子 | 名称: Infectious bursal disease virus mutant HT-VP2-466 / タイプ: sample / ID: 1000 集合状態: 60 copies of IBDV VP2 mutant arranged into 20 trimers Number unique components: 1 |
---|---|
分子量 | 理論値: 2.8 MDa |
-超分子 #1: Infectious bursal disease virus chicken/Cuba/Soroa/1998
超分子 | 名称: Infectious bursal disease virus chicken/Cuba/Soroa/1998 タイプ: virus / ID: 1 / Name.synonym: IBDV / NCBI-ID: 645118 生物種: Infectious bursal disease virus chicken/Cuba/Soroa/1998 ウイルスタイプ: VIRUS-LIKE PARTICLE / ウイルス・単離状態: SPECIES / ウイルス・エンベロープ: No / ウイルス・中空状態: Yes / Syn species name: IBDV |
---|---|
宿主 | 生物種: ![]() ![]() |
Host system | 組換細胞: H5 |
分子量 | 理論値: 2.8 MDa |
ウイルス殻 | Shell ID: 1 / 直径: 700 Å / T番号(三角分割数): 13 |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
---|---|
![]() | 単粒子再構成法 |
試料の集合状態 | particle |
-
試料調製
緩衝液 | pH: 6.2 / 詳細: 25 mM PIPES, 150 mM NaCl, 20 mM CaCl2 |
---|---|
凍結 | 凍結剤: ETHANE / 装置: OTHER |
-
電子顕微鏡法
顕微鏡 | FEI TECNAI F20 |
---|---|
特殊光学系 | エネルギーフィルター - 名称: FEI |
日付 | 2011年4月1日 |
撮影 | カテゴリ: FILM / フィルム・検出器のモデル: KODAK SO-163 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 / 実像数: 88 / 平均電子線量: 10 e/Å2 / ビット/ピクセル: 8 |
電子線 | 加速電圧: 200 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.26 mm / 倍率(公称値): 50000 |
試料ステージ | 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 |
-
画像解析
最終 再構成 | アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 20.8 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: bshow, xmipp / 使用した粒子像数: 1267 |
---|