+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 7fei | ||||||
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タイトル | Complex of FMDV A/WH/CHA/09 and bovine neutralizing scFv antibody R55 | ||||||
要素 |
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キーワード | VIRUS | ||||||
機能・相同性 | 機能・相同性情報 : / RNA-protein covalent cross-linking / : / L-peptidase / icosahedral viral capsid / : / positive stranded viral RNA replication / symbiont-mediated perturbation of host chromatin organization / protein complex oligomerization / ribonucleoside triphosphate phosphatase activity ...: / RNA-protein covalent cross-linking / : / L-peptidase / icosahedral viral capsid / : / positive stranded viral RNA replication / symbiont-mediated perturbation of host chromatin organization / protein complex oligomerization / ribonucleoside triphosphate phosphatase activity / picornain 3C / monoatomic ion channel activity / T=pseudo3 icosahedral viral capsid / host cell cytoplasmic vesicle membrane / endocytosis involved in viral entry into host cell / nucleoside-triphosphate phosphatase / viral capsid / regulation of translation / clathrin-dependent endocytosis of virus by host cell / host cell cytoplasm / RNA helicase activity / viral protein processing / induction by virus of host autophagy / symbiont entry into host cell / RNA-directed RNA polymerase / viral RNA genome replication / cysteine-type endopeptidase activity / RNA-dependent RNA polymerase activity / DNA-templated transcription / virion attachment to host cell / structural molecule activity / RNA binding / ATP binding / membrane / cytoplasm 類似検索 - 分子機能 | ||||||
生物種 | Bos taurus (ウシ) Foot-and-mouth disease virus - type A (ウイルス) | ||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.91 Å | ||||||
データ登録者 | He, Y. / Li, K. / Lou, Z. | ||||||
引用 | ジャーナル: J Virol / 年: 2021 タイトル: Structures of Foot-and-Mouth Disease Virus with Bovine Neutralizing Antibodies Reveal the Determinant of Intraserotype Cross-Neutralization. 著者: Yong He / Kun Li / Li Wang / Zixian Sun / Yimei Cao / Pinghua Li / Pu Sun / Huifang Bao / Shasha Zhou / Sheng Wang / Xingwen Bai / Xuerong Liu / Lixia Zhao / Xiuli Fan / Zaixin Liu / Zengjun ...著者: Yong He / Kun Li / Li Wang / Zixian Sun / Yimei Cao / Pinghua Li / Pu Sun / Huifang Bao / Shasha Zhou / Sheng Wang / Xingwen Bai / Xuerong Liu / Lixia Zhao / Xiuli Fan / Zaixin Liu / Zengjun Lu / Cheng Yang / Zhiyong Lou / 要旨: Foot-and-mouth disease virus (FMDV) exhibits broad antigenic diversity with poor intraserotype cross-neutralizing activity. Studies of the determinant involved in this diversity are essential for the ...Foot-and-mouth disease virus (FMDV) exhibits broad antigenic diversity with poor intraserotype cross-neutralizing activity. Studies of the determinant involved in this diversity are essential for the development of broadly protective vaccines. In this work, we isolated a bovine antibody, designated R55, that displays cross-reaction with both FMDV A/AF/72 (hereafter named FMDV-AAF) and FMDV A/WH/09 (hereafter named FMDV-AWH) but only has a neutralizing effect on FMDV-AWH. Near-atomic resolution structures of FMDV-AAF-R55 and FMDV-AWH-R55 show that R55 engages the capsids of both FMDV-AAF and FMDV-AWH near the icosahedral 3-fold axis and binds to the βB and BC/HI-loops of VP2 and to the B-B knob of VP3. The common interaction residues are highly conserved, which is the major determinant for cross-reaction with both FMDV-AAF and FMDV-AWH. In addition, the cryo-EM structure of the FMDV-AWH-R55 complex also shows that R55 binds to E70 located at the VP3 BC-loop in an adjacent pentamer, which enhances the acid and thermal stabilities of the viral capsid. This may prevent capsid dissociation and genome release into host cells, eventually leading to neutralization of the viral infection. In contrast, R55 binds only to the FMDV-AAF capsid within one pentamer due to the E70G variation, which neither enhances capsid stability nor neutralizes FMDV-AAF infection. The E70G mutation is the major determinant involved in the neutralizing differences between FMDV-AWH and FMDV-AAF. The crucial amino acid E70 is a key component of the neutralizing epitopes, which may aid in the development of broadly protective vaccines. Foot-and-mouth disease virus (FMDV) causes a highly contagious and economically devastating disease in cloven-hoofed animals, and neutralizing antibodies play critical roles in the defense against viral infections. Here, we isolated a bovine antibody (R55) using the single B cell antibody isolation technique. Enzyme-linked immunosorbent assays (ELISA) and virus neutralization tests (VNT) showed that R55 displays cross-reactions with both FMDV-AWH and FMDV-AAF but only has a neutralizing effect on FMDV-AWH. Cryo-EM structures, fluorescence-based thermal stability assays and acid stability assays showed that R55 engages the capsid of FMDV-AWH near the icosahedral 3-fold axis and informs an interpentamer epitope, which overstabilizes virions to hinder capsid dissociation to release the genome, eventually leading to neutralization of viral infection. The crucial amino acid E70 forms a key component of neutralizing epitopes, and the determination of the E70G mutation involved in the neutralizing differences between FMDV-AWH and FMDV-AAF could aid in the development of broadly protective vaccines. | ||||||
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 | 分子: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 7fei.cif.gz | 162.1 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb7fei.ent.gz | 130.9 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 7fei.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 | 7fei_validation.pdf.gz | 913.5 KB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 7fei_full_validation.pdf.gz | 969.8 KB | 表示 | |
XML形式データ | 7fei_validation.xml.gz | 39.7 KB | 表示 | |
CIF形式データ | 7fei_validation.cif.gz | 56.2 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/fe/7fei ftp://data.pdbj.org/pub/pdb/validation_reports/fe/7fei | HTTPS FTP |
-関連構造データ
-リンク
-集合体
登録構造単位 |
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対称性 | 点対称性: (シェーンフリース記号: I (正20面体型対称)) |
-要素
-Capsid protein ... , 4種, 4分子 1234
#1: タンパク質 | 分子量: 23402.678 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) Foot-and-mouth disease virus - type A (ウイルス) 参照: UniProt: E7D639 |
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#2: タンパク質 | 分子量: 24541.584 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) Foot-and-mouth disease virus - type A (ウイルス) 参照: UniProt: J9PFK1, RNA-directed RNA polymerase, nucleoside-triphosphate phosphatase |
#3: タンパク質 | 分子量: 24157.025 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) Foot-and-mouth disease virus - type A (ウイルス) 参照: UniProt: U5JG68 |
#4: タンパク質 | 分子量: 8778.129 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) Foot-and-mouth disease virus - type A (ウイルス) 参照: UniProt: P03309 |
-抗体 , 2種, 2分子 HL
#5: 抗体 | 分子量: 13615.155 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Bos taurus (ウシ) / 発現宿主: Escherichia coli BL21 (大腸菌) |
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#6: 抗体 | 分子量: 12886.850 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Bos taurus (ウシ) / 発現宿主: Escherichia coli BL21 (大腸菌) |
-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
-試料調製
構成要素 |
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由来(天然) |
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由来(組換発現) | 生物種: Escherichia coli BL21 (大腸菌) | ||||||||||||||||||||||||
緩衝液 | pH: 7.4 | ||||||||||||||||||||||||
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||||||||||||||
急速凍結 | 凍結剤: ETHANE |
-電子顕微鏡撮影
実験機器 | モデル: Talos Arctica / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TALOS ARCTICA |
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 200 kV / 照射モード: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELD |
撮影 | 電子線照射量: 25 e/Å2 フィルム・検出器のモデル: DIRECT ELECTRON DE-16 (4k x 4k) |
-解析
CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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3次元再構成 | 解像度: 3.91 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 14535 / 対称性のタイプ: POINT |