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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: PDB / ID: 9ogu | |||||||||||||||||||||||||||||||||||||||||||||
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| タイトル | HIV-1 Env BG505 SOSIP.664-dPG-His in complex with PGT122 and 3BNC117 Fabs | |||||||||||||||||||||||||||||||||||||||||||||
要素 |
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キーワード | VIRAL PROTEIN/Immune System / broadly neutralizing antibody / gp140 / vaccine design / stabilizing mutations / protein design / VIRAL PROTEIN / VIRAL PROTEIN-Immune System complex | |||||||||||||||||||||||||||||||||||||||||||||
| 機能・相同性 | 機能・相同性情報membrane fusion involved in viral entry into host cell / symbiont-mediated perturbation of host defense response / positive regulation of plasma membrane raft polarization / positive regulation of receptor clustering / host cell endosome membrane / clathrin-dependent endocytosis of virus by host cell / viral protein processing / fusion of virus membrane with host plasma membrane / fusion of virus membrane with host endosome membrane / viral envelope ...membrane fusion involved in viral entry into host cell / symbiont-mediated perturbation of host defense response / positive regulation of plasma membrane raft polarization / positive regulation of receptor clustering / host cell endosome membrane / clathrin-dependent endocytosis of virus by host cell / viral protein processing / fusion of virus membrane with host plasma membrane / fusion of virus membrane with host endosome membrane / viral envelope / symbiont entry into host cell / virion attachment to host cell / host cell plasma membrane / virion membrane / structural molecule activity / membrane / identical protein binding 類似検索 - 分子機能 | |||||||||||||||||||||||||||||||||||||||||||||
| 生物種 | ![]() Human immunodeficiency virus 1 (ヒト免疫不全ウイルス) 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: 3.2 Å | |||||||||||||||||||||||||||||||||||||||||||||
データ登録者 | Andrade, T.G. / Ozorowski, G. / Ward, A.B. | |||||||||||||||||||||||||||||||||||||||||||||
| 資金援助 | 米国, 1件
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引用 | ジャーナル: J Virol / 年: 2025タイトル: A modification to heptad repeat 1 of gp41 improves yield and/or quality of soluble pre-fusion HIV-1 envelope glycoprotein trimers. 著者: Devidas N Chaturbhuj / Kwinten Sliepen / Albert Cupo / Benjamin Steinberg / Simon Kazimierczyk / Tarek Munawar / Kyle Kramer / Anila Yasmeen / Thales G Andrade / Wen-Hsin Lee / Lara van der ...著者: Devidas N Chaturbhuj / Kwinten Sliepen / Albert Cupo / Benjamin Steinberg / Simon Kazimierczyk / Tarek Munawar / Kyle Kramer / Anila Yasmeen / Thales G Andrade / Wen-Hsin Lee / Lara van der Maas / Grace Gibson / Oscar Feliciano / Ivan Del Moral Sanchez / Edith Schermer / Rhianna Bronson / Alison Benner / Madhu Prabhakaran / Rosemarie Mason / P J Klasse / Andrew B Ward / Gabriel Ozorowski / Rogier W Sanders / John P Moore / ![]() 要旨: Native-like HIV-1 envelope glycoprotein (Env) trimers, exemplified by the SOSIP design, are widely used as immunogens, analytical antigens, and for structural studies. These vaccine research and ...Native-like HIV-1 envelope glycoprotein (Env) trimers, exemplified by the SOSIP design, are widely used as immunogens, analytical antigens, and for structural studies. These vaccine research and development programs require trimers that are based on multiple HIV-1 genotypes. While a wide range of protein engineering strategies can produce SOSIP trimers from most Env gene sequences, there are still examples of trimers that are expressed only at impractically low yields or that are unstable. Accordingly, additional protein modifications aimed at overcoming such limitations need to be evaluated. Here, we describe a new heptad repeat 1 modification of gp41, known as dPG, that helps to further stabilize the gp41 component of prototypic and germline-targeting SOSIP trimers in the pre-fusion state and thereby increases post-purification yields substantially. The dPG modification involves a deletion (d) at the highly conserved 566 position that disrupts the heptad repeat and introduces proline (P) and glycine (G) substitutions at positions 567 and 568, respectively. We show that the dPG strategy reinforces previously described stabilization changes in existing SOSIP trimers and can rescue otherwise problematic trimer constructs. The latter includes trimers used to target or analyze human germline antibodies and others derived from the global HIV-1 neutralization panel. In summary, the dPG modification strategy can increase the yield and/or quality of Env trimers that are otherwise difficult to produce. IMPORTANCE: Stabilized, soluble, pre-fusion SOSIP trimers are widely used in HIV-1 Env vaccine research. Protein engineering techniques have identified multiple ways to stabilize SOSIP trimers from a ...IMPORTANCE: Stabilized, soluble, pre-fusion SOSIP trimers are widely used in HIV-1 Env vaccine research. Protein engineering techniques have identified multiple ways to stabilize SOSIP trimers from a range of genotypes. However, some SOSIP trimers remain difficult to express at adequate yields and/or purity, so there is a need for additional modifications. Here, we identified a sequence change, designated dPG, to the gp41 subunit that increases the yield and/or quality of various otherwise problematic SOSIP trimers without compromising their antigenicity or structure. This new modification may have general value for HIV-1 vaccine research and development. | |||||||||||||||||||||||||||||||||||||||||||||
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構造の表示
| 構造ビューア | 分子: Molmil Jmol/JSmol |
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ダウンロードとリンク
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ダウンロード
| PDBx/mmCIF形式 | 9ogu.cif.gz | 668.6 KB | 表示 | PDBx/mmCIF形式 |
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| PDB形式 | pdb9ogu.ent.gz | 535.6 KB | 表示 | PDB形式 |
| PDBx/mmJSON形式 | 9ogu.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
| アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/og/9ogu ftp://data.pdbj.org/pub/pdb/validation_reports/og/9ogu | HTTPS FTP |
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-関連構造データ
| 関連構造データ | ![]() 70476MC ![]() 9ogtC M: このデータのモデリングに利用したマップデータ C: 同じ文献を引用 ( |
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| 類似構造データ | 類似検索 - 機能・相同性 F&H 検索 |
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リンク
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集合体
| 登録構造単位 | ![]()
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要素
-タンパク質 , 2種, 6分子 AECFDB
| #1: タンパク質 | 分子量: 57945.977 Da / 分子数: 3 / 断片: UNP residues 30-505 / 変異: T332N, A501C / 由来タイプ: 組換発現 由来: (組換発現) ![]() Human immunodeficiency virus 1 (ヒト免疫不全ウイルス)遺伝子: env / 発現宿主: Homo sapiens (ヒト) / 参照: UniProt: Q2N0S6#2: タンパク質 | 分子量: 18596.820 Da / 分子数: 3 / 変異: K567P, L568G / 由来タイプ: 組換発現 由来: (組換発現) ![]() Human immunodeficiency virus 1 (ヒト免疫不全ウイルス)遺伝子: env / 発現宿主: Homo sapiens (ヒト) / 参照: UniProt: A0A6H1VYE9 |
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-抗体 , 4種, 12分子 HMPILQJNRKOS
| #3: 抗体 | 分子量: 24656.484 Da / 分子数: 3 / 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: 抗体 | 分子量: 23022.658 Da / 分子数: 3 / 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)#5: 抗体 | 分子量: 25434.691 Da / 分子数: 3 / 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)#6: 抗体 | 分子量: 22712.082 Da / 分子数: 3 / 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) |
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-糖 , 9種, 54分子 
| #7: 多糖 | 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose #8: 多糖 | alpha-D-mannopyranose-(1-3)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1- ...alpha-D-mannopyranose-(1-3)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose | #9: 多糖 | alpha-D-mannopyranose-(1-2)-alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D- ...alpha-D-mannopyranose-(1-2)-alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose | #10: 多糖 | #11: 多糖 | alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2- ...alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose #12: 多糖 | beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta- ...beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose #13: 多糖 | #14: 多糖 | beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose | #15: 糖 | ChemComp-NAG / |
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-詳細
| 研究の焦点であるリガンドがあるか | N |
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| Has protein modification | Y |
-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 |
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試料調製
| 構成要素 | 名称: HIV-1 Env BG505 SOSIP.664-dPG-His in complex with PGT122 and 3BNC117 Fabs タイプ: COMPLEX / Entity ID: #1-#6 / 由来: RECOMBINANT |
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| 由来(天然) | 生物種: ![]() Human immunodeficiency virus 1 (ヒト免疫不全ウイルス) |
| 由来(組換発現) | 生物種: Homo sapiens (ヒト) |
| 緩衝液 | pH: 7.4 |
| 試料 | 濃度: 2.6 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
| 試料支持 | グリッドの材料: GOLD / グリッドのサイズ: 300 divisions/in. / グリッドのタイプ: UltrAuFoil R1.2/1.3 |
| 急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 277 K |
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電子顕微鏡撮影
| 顕微鏡 | モデル: TFS GLACIOS |
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| 電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 200 kV / 照射モード: FLOOD BEAM |
| 電子レンズ | モード: BRIGHT FIELD / 倍率(公称値): 190000 X / 最大 デフォーカス(公称値): 1800 nm / 最小 デフォーカス(公称値): 800 nm |
| 撮影 | 電子線照射量: 45 e/Å2 フィルム・検出器のモデル: FEI FALCON IV (4k x 4k) |
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解析
| EMソフトウェア |
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| CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
| 対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||||||
| 3次元再構成 | 解像度: 3.2 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 210222 / 対称性のタイプ: POINT | ||||||||||||||||||||||||
| 精密化 | 最高解像度: 3.2 Å 立体化学のターゲット値: REAL-SPACE (WEIGHTED MAP SUM AT ATOM CENTERS) | ||||||||||||||||||||||||
| 拘束条件 |
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万見について





Human immunodeficiency virus 1 (ヒト免疫不全ウイルス)
Homo sapiens (ヒト)
米国, 1件
引用



PDBj





FIELD EMISSION GUN