ジャーナル: Immunity / 年: 2015 タイトル: Affinity Maturation of a Potent Family of HIV Antibodies Is Primarily Focused on Accommodating or Avoiding Glycans. 著者: Fernando Garces / Jeong Hyun Lee / Natalia de Val / Alba Torrents de la Pena / Leopold Kong / Cristina Puchades / Yuanzi Hua / Robyn L Stanfield / Dennis R Burton / John P Moore / Rogier W ...著者: Fernando Garces / Jeong Hyun Lee / Natalia de Val / Alba Torrents de la Pena / Leopold Kong / Cristina Puchades / Yuanzi Hua / Robyn L Stanfield / Dennis R Burton / John P Moore / Rogier W Sanders / Andrew B Ward / Ian A Wilson / 要旨: The high-mannose patch on the HIV-1 envelope (Env) glycoprotein is the epicenter for binding of the potent broadly neutralizing PGT121 family of antibodies, but strategies for generating such ...The high-mannose patch on the HIV-1 envelope (Env) glycoprotein is the epicenter for binding of the potent broadly neutralizing PGT121 family of antibodies, but strategies for generating such antibodies by vaccination have not been defined. We generated structures of inferred antibody intermediates by X-ray crystallography and electron microscopy to elucidate the molecular events that occurred during evolution of this family. Binding analyses revealed that affinity maturation was primarily focused on avoiding, accommodating, or binding the N137 glycan. The overall antibody approach angle to Env was defined very early in the maturation process, yet some variation evolved in the PGT121 family branches that led to differences in glycan specificities in their respective epitopes. Furthermore, we determined a crystal structure of the recombinant BG505 SOSIP.664 HIV-1 trimer with a PGT121 family member at 3.0 Å that, in concert with these antibody intermediate structures, provides insights to advance design of HIV vaccine candidates.
生物種: Human Immunodeficiency Virus-1 (ヒト免疫不全ウイルス) 別称: HIV-1
分子量
理論値: 420 MDa
組換発現
生物種: Mammalian (両生類) / 組換細胞: HEK293S
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分子 #2: PGT124 Fab with 32H heavy chain
分子
名称: PGT124 Fab with 32H heavy chain / タイプ: protein_or_peptide / ID: 2 / Name.synonym: PGT124 32H-3L 詳細: One light and one heavy chain forms the Fab heterodimer コピー数: 3 / 集合状態: Heterodimer / 組換発現: Yes
由来(天然)
生物種: Homo sapiens (ヒト) / 別称: Human
分子量
理論値: 50 KDa
組換発現
生物種: Homo sapiens (ヒト) / 組換細胞: HEK293F
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実験情報
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構造解析
手法
ネガティブ染色法
解析
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
緩衝液
pH: 7.4 / 詳細: 20 mM Tris, 150 mM NaCl
染色
タイプ: NEGATIVE 詳細: Grids adsorbed with protein then stained with 2% w/v uranyl formate
グリッド
詳細: 400 mesh Cu grid with carbon support, plasma cleaned