ジャーナル: PLoS Pathog / 年: 2015 タイトル: Comprehensive antigenic map of a cleaved soluble HIV-1 envelope trimer. 著者: Ronald Derking / Gabriel Ozorowski / Kwinten Sliepen / Anila Yasmeen / Albert Cupo / Jonathan L Torres / Jean-Philippe Julien / Jeong Hyun Lee / Thijs van Montfort / Steven W de Taeye / Mark ...著者: Ronald Derking / Gabriel Ozorowski / Kwinten Sliepen / Anila Yasmeen / Albert Cupo / Jonathan L Torres / Jean-Philippe Julien / Jeong Hyun Lee / Thijs van Montfort / Steven W de Taeye / Mark Connors / Dennis R Burton / Ian A Wilson / Per-Johan Klasse / Andrew B Ward / John P Moore / Rogier W Sanders / 要旨: The trimeric envelope (Env) spike is the focus of vaccine design efforts aimed at generating broadly neutralizing antibodies (bNAbs) to protect against HIV-1 infection. Three recent developments have ...The trimeric envelope (Env) spike is the focus of vaccine design efforts aimed at generating broadly neutralizing antibodies (bNAbs) to protect against HIV-1 infection. Three recent developments have facilitated a thorough investigation of the antigenic structure of the Env trimer: 1) the isolation of many bNAbs against multiple different epitopes; 2) the generation of a soluble trimer mimic, BG505 SOSIP.664 gp140, that expresses most bNAb epitopes; 3) facile binding assays involving the oriented immobilization of tagged trimers. Using these tools, we generated an antigenic map of the trimer by antibody cross-competition. Our analysis delineates three well-defined epitope clusters (CD4 binding site, quaternary V1V2 and Asn332-centered oligomannose patch) and new epitopes at the gp120-gp41 interface. It also identifies the relationships among these clusters. In addition to epitope overlap, we defined three more ways in which antibodies can cross-compete: steric competition from binding to proximal but non-overlapping epitopes (e.g., PGT151 inhibition of 8ANC195 binding); allosteric inhibition (e.g., PGT145 inhibition of 1NC9, 8ANC195, PGT151 and CD4 binding); and competition by reorientation of glycans (e.g., PGT135 inhibition of CD4bs bNAbs, and CD4bs bNAb inhibition of 8ANC195). We further demonstrate that bNAb binding can be complex, often affecting several other areas of the trimer surface beyond the epitope. This extensive analysis of the antigenic structure and the epitope interrelationships of the Env trimer should aid in design of both bNAb-based therapies and vaccines intended to induce bNAbs.
全体 : Fab of 1NC9 human monoclonal antibody bound to HIV-1 Env gp140 BG...
全体
名称: Fab of 1NC9 human monoclonal antibody bound to HIV-1 Env gp140 BG505 SOSIP.664
要素
試料: Fab of 1NC9 human monoclonal antibody bound to HIV-1 Env gp140 BG505 SOSIP.664
タンパク質・ペプチド: Soluble HIV-1 Envelope glycoprotein
タンパク質・ペプチド: Fab of 1NC9 human monoclonal antibody
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超分子 #1000: Fab of 1NC9 human monoclonal antibody bound to HIV-1 Env gp140 BG...
超分子
名称: Fab of 1NC9 human monoclonal antibody bound to HIV-1 Env gp140 BG505 SOSIP.664 タイプ: sample / ID: 1000 詳細: Size-exclusion-chromatography-purified gp140 trimers were used. 集合状態: One Fab molecule binds to one BG505 SOSIP.664 gp140 monomer Number unique components: 2
試料ホルダーモデル: SIDE ENTRY, EUCENTRIC / Tilt angle min: -50
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画像解析
詳細
The particles were selected using an automatic selection program. Class averages with 3 Fabs bound to one gp140 trimer were placed into a substack and used for 3D reconstruction.
最終 再構成
アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 18.0 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: EMAN2, SPARX / 使用した粒子像数: 11235