+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 3iyw | |||||||||||||||
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タイトル | West Nile virus in complex with Fab fragments of MAb CR4354 (fitted coordinates of envelope proteins and Fab fragments of one icosahedral ASU) | |||||||||||||||
要素 |
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キーワード | VIRUS / virus-antibody complex / neutralizing Fab fragment / flavivirus / West Nile Virus / envelope protein / icosahedral virus | |||||||||||||||
機能・相同性 | 機能・相同性情報 flavivirin / symbiont-mediated suppression of host JAK-STAT cascade via inhibition of STAT2 activity / symbiont-mediated suppression of host JAK-STAT cascade via inhibition of STAT1 activity / double-stranded RNA binding / viral capsid / nucleoside-triphosphate phosphatase / mRNA (guanine-N7)-methyltransferase / methyltransferase cap1 / mRNA (nucleoside-2'-O-)-methyltransferase activity / mRNA 5'-cap (guanine-N7-)-methyltransferase activity ...flavivirin / symbiont-mediated suppression of host JAK-STAT cascade via inhibition of STAT2 activity / symbiont-mediated suppression of host JAK-STAT cascade via inhibition of STAT1 activity / double-stranded RNA binding / viral capsid / nucleoside-triphosphate phosphatase / mRNA (guanine-N7)-methyltransferase / methyltransferase cap1 / mRNA (nucleoside-2'-O-)-methyltransferase activity / mRNA 5'-cap (guanine-N7-)-methyltransferase activity / RNA helicase activity / host cell endoplasmic reticulum membrane / host cell perinuclear region of cytoplasm / protein dimerization activity / symbiont-mediated suppression of host type I interferon-mediated signaling pathway / RNA helicase / symbiont entry into host cell / 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 | West Nile virus (西ナイルウイルス) Homo sapiens (ヒト) | |||||||||||||||
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: 13.7 Å | |||||||||||||||
データ登録者 | Rossmann, M.G. / Kaufmann, B. | |||||||||||||||
引用 | ジャーナル: Proc Natl Acad Sci U S A / 年: 2010 タイトル: Neutralization of West Nile virus by cross-linking of its surface proteins with Fab fragments of the human monoclonal antibody CR4354. 著者: Bärbel Kaufmann / Matthew R Vogt / Jaap Goudsmit / Heather A Holdaway / Anastasia A Aksyuk / Paul R Chipman / Richard J Kuhn / Michael S Diamond / Michael G Rossmann / 要旨: Many flaviviruses are significant human pathogens, with the humoral immune response playing an essential role in restricting infection and disease. CR4354, a human monoclonal antibody isolated from a ...Many flaviviruses are significant human pathogens, with the humoral immune response playing an essential role in restricting infection and disease. CR4354, a human monoclonal antibody isolated from a patient, neutralizes West Nile virus (WNV) infection at a postattachment stage in the viral life-cycle. Here, we determined the structure of WNV complexed with Fab fragments of CR4354 using cryoelectron microscopy. The outer glycoprotein shell of a mature WNV particle is formed by 30 rafts of three homodimers of the viral surface protein E. CR4354 binds to a discontinuous epitope formed by protein segments from two neighboring E molecules, but does not cause any detectable structural disturbance on the viral surface. The epitope occurs at two independent positions within an icosahedral asymmetric unit, resulting in 120 binding sites on the viral surface. The cross-linking of the six E monomers within one raft by four CR4354 Fab fragments suggests that the antibody neutralizes WNV by blocking the pH-induced rearrangement of the E protein required for virus fusion with the endosomal membrane. | |||||||||||||||
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 | 3iyw.cif.gz | 409.2 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb3iyw.ent.gz | 332.8 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 3iyw.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 | 3iyw_validation.pdf.gz | 983.6 KB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 3iyw_full_validation.pdf.gz | 1.1 MB | 表示 | |
XML形式データ | 3iyw_validation.xml.gz | 77.3 KB | 表示 | |
CIF形式データ | 3iyw_validation.cif.gz | 115 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/iy/3iyw ftp://data.pdbj.org/pub/pdb/validation_reports/iy/3iyw | 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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1 |
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2 |
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3 |
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5 |
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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
#1: タンパク質 | 分子量: 43272.098 Da / 分子数: 3 / 断片: ectodomain of viral surface protein / 由来タイプ: 組換発現 由来: (組換発現) West Nile virus (西ナイルウイルス) 発現宿主: Homo sapiens (ヒト) / 参照: UniProt: Q91R02, UniProt: Q9Q6P4*PLUS #2: 抗体 | 分子量: 24745.699 Da / 分子数: 2 / 断片: Fab fragment, heavy chain / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / Cell (発現宿主): B cells / 発現宿主: Homo sapiens (ヒト) / 組織 (発現宿主): peripheral blood #3: 抗体 | 分子量: 22908.160 Da / 分子数: 2 / 断片: Fab fragment, light chain / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 発現宿主: Homo sapiens (ヒト) #4: 多糖 | |
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-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 | 名称: West Nile virus in complex with Fab fragments of MAb CR4354 タイプ: VIRUS 詳細: THE ENTRY PRESENTED HERE DOES NOT CONTAIN THE COMPLETE BIOLOGICAL ASSEMBLY. COORDINATES FOR A COMPLETE MULTIMER REPRESENTING THE KNOWN BIOLOGICALLY SIGNIFICANT OLIGOMERIZATION STATE OF THE ...詳細: THE ENTRY PRESENTED HERE DOES NOT CONTAIN THE COMPLETE BIOLOGICAL ASSEMBLY. COORDINATES FOR A COMPLETE MULTIMER REPRESENTING THE KNOWN BIOLOGICALLY SIGNIFICANT OLIGOMERIZATION STATE OF THE VIRUS-FAB COMPLEX CAN BE GENERATED BY APPLYING BIOMT TRANSFORMATIONS GIVEN. (entry represents one icoshedral ASU containing 3 envelope E protein molecules and 2 CR4354 Fab fragments) |
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ウイルスについての詳細 | 中空か: NO / エンベロープを持つか: YES / ホストのカテゴリ: VERTEBRATES / 単離: STRAIN / タイプ: VIRION |
天然宿主 | 生物種: Homo sapiens |
緩衝液 | pH: 8 / 詳細: 12mM Tris-HCl, 120mM NaCl, 1mM EDTA |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES / 詳細: 12mM Tris-HCl, 120mM NaCl, 1mM EDTA |
急速凍結 | 装置: HOMEMADE PLUNGER / 凍結剤: ETHANE 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. |
-電子顕微鏡撮影
顕微鏡 | モデル: FEI/PHILIPS CM300FEG/T / 日付: 2009年9月9日 / 詳細: low dose imaging |
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電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELD / 倍率(公称値): 45000 X / 倍率(補正後): 47244 X / 最大 デフォーカス(公称値): 3530 nm / 最小 デフォーカス(公称値): 1450 nm / 非点収差: live FFT at 200K magnification / カメラ長: 0 mm |
試料ホルダ | 試料ホルダーモデル: GATAN LIQUID NITROGEN / 資料ホルダタイプ: Eucentric / 温度: 98 K / 傾斜角・最大: -9999 ° / 傾斜角・最小: -9999 ° |
撮影 | 電子線照射量: 22 e/Å2 / フィルム・検出器のモデル: KODAK SO-163 FILM |
画像スキャン | サンプリングサイズ: 6.35 µm / 詳細: scanned images binned 2x2 / デジタル画像の数: 69 / Od range: 1 / Scanner model: NIKON SUPER COOLSCAN 9000 |
放射 | プロトコル: SINGLE WAVELENGTH / 単色(M)・ラウエ(L): M |
放射波長 | 相対比: 1 |
-解析
EMソフトウェア |
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CTF補正 | 詳細: Each particle | |||||||||||||||||||||
対称性 | 点対称性: I (正20面体型対称) | |||||||||||||||||||||
3次元再構成 | 手法: common lines, Fourier method / 解像度: 13.7 Å / 解像度の算出法: FSC 0.5 CUT-OFF / 粒子像の数: 5006 詳細: final map includes data to 13.0 Ang resolution (FCS at about 0.2 cut-off) ( Details about the particle: The particles were selected interactively at the computer terminal. ) 対称性のタイプ: POINT | |||||||||||||||||||||
原子モデル構築 |
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原子モデル構築 |
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精密化ステップ | サイクル: LAST
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