National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
UM1AI100663
米国
Bill & Melinda Gates Foundation
OPP1115782
米国
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
UM1AI144462
米国
引用
ジャーナル: Cell Rep / 年: 2020 タイトル: HIV-1 Envelope and MPER Antibody Structures in Lipid Assemblies. 著者: Kimmo Rantalainen / Zachary T Berndsen / Aleksandar Antanasijevic / Torben Schiffner / Xi Zhang / Wen-Hsin Lee / Jonathan L Torres / Lei Zhang / Adriana Irimia / Jeffrey Copps / Kenneth H ...著者: Kimmo Rantalainen / Zachary T Berndsen / Aleksandar Antanasijevic / Torben Schiffner / Xi Zhang / Wen-Hsin Lee / Jonathan L Torres / Lei Zhang / Adriana Irimia / Jeffrey Copps / Kenneth H Zhou / Young D Kwon / William H Law / Chaim A Schramm / Raffaello Verardi / Shelly J Krebs / Peter D Kwong / Nicole A Doria-Rose / Ian A Wilson / Michael B Zwick / John R Yates / William R Schief / Andrew B Ward / 要旨: Structural and functional studies of HIV envelope glycoprotein (Env) as a transmembrane protein have long been complicated by challenges associated with inherent flexibility of the molecule and the ...Structural and functional studies of HIV envelope glycoprotein (Env) as a transmembrane protein have long been complicated by challenges associated with inherent flexibility of the molecule and the membrane-embedded hydrophobic regions. Here, we present approaches for incorporating full-length, wild-type HIV-1 Env, as well as C-terminally truncated and stabilized versions, into lipid assemblies, providing a modular platform for Env structural studies by single particle electron microscopy. We reconstitute a full-length Env clone into a nanodisc, complex it with a membrane-proximal external region (MPER) targeting antibody 10E8, and structurally define the full quaternary epitope of 10E8 consisting of lipid, MPER, and ectodomain contacts. By aligning this and other Env-MPER antibody complex reconstructions with the lipid bilayer, we observe evidence of Env tilting as part of the neutralization mechanism for MPER-targeting antibodies. We also adapt the platform toward vaccine design purposes by introducing stabilizing mutations that allow purification of unliganded Env with a peptidisc scaffold.
名称: Full-length HIV-1 Envelope glycoprotein clone AMC011 in complex with PGT151 Fab and 10E8v4-5R+100cF Fab タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#11 詳細: AMC011 Env purified with stabilizing PGT151 Fab. Complexed with 10E8v4-5R+100cF Fab during detergent-lipid exchange and prior to grid preparation.
由来(天然)
生物種: Homo sapiens (ヒト)
組換発現
生物種: Homo sapiens (ヒト)
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実験情報
-
構造解析
手法
クライオ電子顕微鏡法
解析
単粒子再構成法
試料の集合状態
particle
-
試料調製
濃度
5.2 mg/mL
緩衝液
pH: 7.4
グリッド
モデル: C-flat-1.2/1.3 / 材質: COPPER / メッシュ: 400
凍結
凍結剤: ETHANE / チャンバー内湿度: 100 % / 装置: FEI VITROBOT MARK IV
詳細
AMC011 Env purified with stabilizing PGT151 Fab. Complexed with 10E8v4-5R+100cF Fab during detergent-lipid exchange and prior to grid preparation. Sample is in detergent-lipid micelle.