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データを開く
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基本情報
登録情報 | データベース: EMDB / ID: EMD-3086 | |||||||||
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タイトル | negative stain EM of BG505 SOSIP.664 in complex with sCD4, 17b, and 8ANC195 | |||||||||
![]() | negative stain EM single particle analysis. The particles were picked using EMAN2. CTF correction was done using EMAN2. 2D and 3D classification was done using Relion to sort particles. The refinement and gold-standard FSC calculation was done using Relion. | |||||||||
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![]() | HIV-1 Env trimer / broadly neutralizing antibody | |||||||||
機能・相同性 | ![]() helper T cell enhancement of adaptive immune response / interleukin-16 binding / interleukin-16 receptor activity / maintenance of protein location in cell / T cell selection / MHC class II protein binding / cellular response to granulocyte macrophage colony-stimulating factor stimulus / interleukin-15-mediated signaling pathway / positive regulation of monocyte differentiation / Nef Mediated CD4 Down-regulation ...helper T cell enhancement of adaptive immune response / interleukin-16 binding / interleukin-16 receptor activity / maintenance of protein location in cell / T cell selection / MHC class II protein binding / cellular response to granulocyte macrophage colony-stimulating factor stimulus / interleukin-15-mediated signaling pathway / positive regulation of monocyte differentiation / Nef Mediated CD4 Down-regulation / Alpha-defensins / positive regulation of kinase activity / regulation of T cell activation / T cell receptor complex / extracellular matrix structural constituent / Other interleukin signaling / enzyme-linked receptor protein signaling pathway / Dectin-2 family / Translocation of ZAP-70 to Immunological synapse / Phosphorylation of CD3 and TCR zeta chains / regulation of calcium ion transport / macrophage differentiation / Generation of second messenger molecules / T cell differentiation / PD-1 signaling / positive regulation of protein kinase activity / Binding and entry of HIV virion / coreceptor activity / positive regulation of calcium-mediated signaling / positive regulation of plasma membrane raft polarization / positive regulation of receptor clustering / cell surface receptor protein tyrosine kinase signaling pathway / positive regulation of establishment of T cell polarity / T cell activation / positive regulation of interleukin-2 production / protein tyrosine kinase binding / host cell endosome membrane / Vpu mediated degradation of CD4 / calcium-mediated signaling / clathrin-coated endocytic vesicle membrane / transmembrane signaling receptor activity / positive regulation of peptidyl-tyrosine phosphorylation / Downstream TCR signaling / Cargo recognition for clathrin-mediated endocytosis / positive regulation of T cell activation / MHC class II protein complex binding / Clathrin-mediated endocytosis / virus receptor activity / signaling receptor activity / clathrin-dependent endocytosis of virus by host cell / positive regulation of canonical NF-kappaB signal transduction / defense response to Gram-negative bacterium / adaptive immune response / positive regulation of MAPK cascade / positive regulation of viral entry into host cell / cell surface receptor signaling pathway / positive regulation of ERK1 and ERK2 cascade / early endosome / viral protein processing / cell adhesion / positive regulation of protein phosphorylation / immune response / membrane raft / endoplasmic reticulum lumen / fusion of virus membrane with host plasma membrane / external side of plasma membrane / virus-mediated perturbation of host defense response / fusion of virus membrane with host endosome membrane / viral envelope / lipid binding / virion attachment to host cell / endoplasmic reticulum membrane / protein kinase binding / host cell plasma membrane / structural molecule activity / virion membrane / positive regulation of DNA-templated transcription / enzyme binding / signal transduction / protein homodimerization activity / zinc ion binding / identical protein binding / membrane / plasma membrane 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / ネガティブ染色法 / 解像度: 16.8 Å | |||||||||
![]() | Scharf L / Wang H / Gao H / Chen S / McDowall A / Bjorkman P | |||||||||
![]() | ![]() タイトル: Broadly Neutralizing Antibody 8ANC195 Recognizes Closed and Open States of HIV-1 Env. 著者: Louise Scharf / Haoqing Wang / Han Gao / Songye Chen / Alasdair W McDowall / Pamela J Bjorkman / ![]() 要旨: The HIV-1 envelope (Env) spike contains limited epitopes for broadly neutralizing antibodies (bNAbs); thus, most neutralizing antibodies are strain specific. The 8ANC195 epitope, defined by crystal ...The HIV-1 envelope (Env) spike contains limited epitopes for broadly neutralizing antibodies (bNAbs); thus, most neutralizing antibodies are strain specific. The 8ANC195 epitope, defined by crystal and electron microscopy (EM) structures of bNAb 8ANC195 complexed with monomeric gp120 and trimeric Env, respectively, spans the gp120 and gp41 Env subunits. To investigate 8ANC195's gp41 epitope at higher resolution, we solved a 3.58 Å crystal structure of 8ANC195 complexed with fully glycosylated Env trimer, revealing 8ANC195 insertion into a glycan shield gap to contact gp120 and gp41 glycans and protein residues. To determine whether 8ANC195 recognizes the CD4-bound open Env conformation that leads to co-receptor binding and fusion, one of several known conformations of virion-associated Env, we solved EM structures of an Env/CD4/CD4-induced antibody/8ANC195 complex. 8ANC195 binding partially closed the CD4-bound trimer, confirming structural plasticity of Env by revealing a previously unseen conformation. 8ANC195's ability to bind different Env conformations suggests advantages for potential therapeutic applications. | |||||||||
履歴 |
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構造の表示
ムービー |
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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ダウンロードとリンク
-EMDBアーカイブ
マップデータ | ![]() | 6 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 14.2 KB 14.2 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 4.9 KB | 表示 | ![]() |
画像 | ![]() | 1.1 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 261.7 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 260.8 KB | 表示 | |
XML形式データ | ![]() | 7.3 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
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リンク
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | negative stain EM single particle analysis. The particles were picked using EMAN2. CTF correction was done using EMAN2. 2D and 3D classification was done using Relion to sort particles. The refinement and gold-standard FSC calculation was done using Relion. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 2.5 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
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試料の構成要素
-全体 : soluble HIV-1 Env trimer BG505 SOSIP.664 in complex with soluble ...
全体 | 名称: soluble HIV-1 Env trimer BG505 SOSIP.664 in complex with soluble CD4s (D1-D2), broadly neutralizing antibody 17b Fabs, and broadly neutralizing antibody 8ANC195 variant G32K5 Fabs |
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要素 |
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-超分子 #1000: soluble HIV-1 Env trimer BG505 SOSIP.664 in complex with soluble ...
超分子 | 名称: soluble HIV-1 Env trimer BG505 SOSIP.664 in complex with soluble CD4s (D1-D2), broadly neutralizing antibody 17b Fabs, and broadly neutralizing antibody 8ANC195 variant G32K5 Fabs タイプ: sample / ID: 1000 集合状態: 3 Fabs of 17b, 3 sCD4, and 3 Fabs of 8ANC195 G52K5 bind to Env trimer BG505 SOSIP Number unique components: 4 |
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分子量 | 理論値: 500 KDa |
-分子 #1: Soluble HIV-1 Env trimer BG505 SOSIP.664
分子 | 名称: Soluble HIV-1 Env trimer BG505 SOSIP.664 / タイプ: protein_or_peptide / ID: 1 / Name.synonym: BG505 SOSIP / コピー数: 1 / 集合状態: trimer / 組換発現: Yes |
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由来(天然) | 生物種: ![]() ![]() 別称: HIV-1 |
分子量 | 理論値: 420 KDa |
組換発現 | 生物種: ![]() |
-分子 #2: Fab of broadly neutralizing antibody 17b
分子 | 名称: Fab of broadly neutralizing antibody 17b / タイプ: protein_or_peptide / ID: 2 / Name.synonym: 17b / コピー数: 3 / 集合状態: monomer / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 50 KDa |
組換発現 | 生物種: ![]() |
-分子 #3: soluble CD4 (D1-D2 domains)
分子 | 名称: soluble CD4 (D1-D2 domains) / タイプ: protein_or_peptide / ID: 3 / Name.synonym: sCD4 / コピー数: 3 / 集合状態: monomer / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 20 KDa |
組換発現 | 生物種: ![]() |
-分子 #4: Fab of broadly neutralizing antibody 8ANC195 variant G52K5
分子 | 名称: Fab of broadly neutralizing antibody 8ANC195 variant G52K5 タイプ: protein_or_peptide / ID: 4 / Name.synonym: 8ANC195 G52K5 / コピー数: 3 / 集合状態: monomer / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 50 KDa |
組換発現 | 生物種: ![]() |
-実験情報
-構造解析
手法 | ネガティブ染色法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
濃度 | 0.01 mg/mL |
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緩衝液 | pH: 8 / 詳細: 20mM Tris, 50mM NaCl |
染色 | タイプ: NEGATIVE 詳細: Grids with adsorbed protein cross-linked by glutaraldehyde vapor and stained by 3% w/v uranyl acetate for 30 seconds. |
グリッド | 詳細: glow discharged ultrathin C film on holey carbon support film, 400 mesh, Cu grids |
凍結 | 凍結剤: NONE / 装置: OTHER |
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電子顕微鏡法
顕微鏡 | FEI TECNAI 12 |
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日付 | 2015年4月7日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: GATAN ULTRASCAN 1000 (2k x 2k) 実像数: 642 |
電子線 | 加速電圧: 120 kV / 電子線源: LAB6 |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.2 mm / 倍率(公称値): 42000 |
試料ステージ | 試料ホルダーモデル: SIDE ENTRY, EUCENTRIC |
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画像解析
-原子モデル構築 1
初期モデル | PDB ID: Chain - #0 - Chain ID: G / Chain - #1 - Chain ID: C / Chain - #2 - Chain ID: L / Chain - #3 - Chain ID: H |
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ソフトウェア | 名称: ![]() |
詳細 | Fitted by manual docking first and then using Chimera with the following options: real-time correlation/average update, and use map simulated from atoms, resolution 15 A |
精密化 | 空間: REAL / プロトコル: RIGID BODY FIT |
得られたモデル | ![]() PDB-5a7x: |
-原子モデル構築 2
初期モデル | PDB ID: Chain - #0 - Chain ID: L / Chain - #1 - Chain ID: H |
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ソフトウェア | 名称: ![]() |
詳細 | Fitted by manual docking first and then using Chimera with the following options: real-time correlation/average update, and use map simulated from atoms, resolution 15 A |
精密化 | 空間: REAL / プロトコル: RIGID BODY FIT |
得られたモデル | ![]() PDB-5a7x: |