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基本情報
登録情報 | データベース: PDB / ID: 5fuu | |||||||||
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タイトル | Ectodomain of cleaved wild type JR-FL EnvdCT trimer in complex with PGT151 Fab | |||||||||
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![]() | VIRAL PROTEIN / HIV-1 / ENV / PGT151 / BROADLY NEUTRALIZING ANTIBODY | |||||||||
機能・相同性 | ![]() 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 / virion attachment to host cell / host cell plasma membrane ...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 / virion attachment to host cell / host cell plasma membrane / virion membrane / structural molecule activity / identical protein binding / membrane 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() ![]() | |||||||||
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: 4.19 Å | |||||||||
![]() | Lee, J.H. / Ward, A.B. | |||||||||
![]() | ![]() タイトル: Cryo-EM structure of a native, fully glycosylated, cleaved HIV-1 envelope trimer. 著者: Jeong Hyun Lee / Gabriel Ozorowski / Andrew B Ward / ![]() 要旨: The envelope glycoprotein trimer (Env) on the surface of HIV-1 recognizes CD4(+) T cells and mediates viral entry. During this process, Env undergoes substantial conformational rearrangements, making ...The envelope glycoprotein trimer (Env) on the surface of HIV-1 recognizes CD4(+) T cells and mediates viral entry. During this process, Env undergoes substantial conformational rearrangements, making it difficult to study in its native state. Soluble stabilized trimers have provided valuable insights into the Env structure, but they lack the hydrophobic membrane proximal external region (MPER, an important target of broadly neutralizing antibodies), the transmembrane domain, and the cytoplasmic tail. Here we present (i) a cryogenic electron microscopy (cryo-EM) structure of a clade B virus Env, which lacks only the cytoplasmic tail and is stabilized by the broadly neutralizing antibody PGT151, at a resolution of 4.2 angstroms and (ii) a reconstruction of this form of Env in complex with PGT151 and MPER-targeting antibody 10E8 at a resolution of 8.8 angstroms. These structures provide new insights into the wild-type Env structure. | |||||||||
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構造の表示
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構造ビューア | 分子: ![]() ![]() |
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-検証レポート
文書・要旨 | ![]() | 4.4 MB | 表示 | ![]() |
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文書・詳細版 | ![]() | 4.5 MB | 表示 | |
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CIF形式データ | ![]() | 106.2 KB | 表示 | |
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-関連構造データ
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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
-HIV-1 ENVELOPE GLYCOPROTEIN ... , 2種, 6分子 ACEBDF
#1: タンパク質 | 分子量: 53342.355 Da / 分子数: 3 / 断片: GP120, RESIDUES 30-502 / 変異: YES / 由来タイプ: 組換発現 / 詳細: GP120 OF JR-FL ENV 由来: (組換発現) ![]() ![]() 遺伝子: ENV / プラスミド: PHCMV3 / 細胞株 (発現宿主): HEK293 / 発現宿主: ![]() #2: タンパク質 | 分子量: 17243.592 Da / 分子数: 3 / 断片: GP41, RESIDUES 503-655 / 由来タイプ: 組換発現 / 詳細: GP41 ECTODOMAIN OF JR-FL ENV 由来: (組換発現) ![]() ![]() 遺伝子: ENV / プラスミド: PHCMV3 / 細胞株 (発現宿主): HEK293 / 発現宿主: ![]() |
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-抗体 , 2種, 4分子 HMLN
#3: 抗体 | 分子量: 26087.438 Da / 分子数: 2 / 断片: FAB HEAVY CHAIN VARIABLE REGION, RESIDUES 1-218 / 由来タイプ: 組換発現 / 詳細: PGT151 CLEAVED INTO FAB / 由来: (組換発現) ![]() ![]() #4: 抗体 | 分子量: 24057.809 Da / 分子数: 2 / 断片: FAB LIGHT CHAIN VARIABLE REGION, RESIDUES 1-214 / 由来タイプ: 組換発現 / 詳細: PGT151 CLEAVED INTO FAB / 由来: (組換発現) ![]() ![]() |
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-糖 , 12種, 64分子 
#5: 多糖 | 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose #6: 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 #7: 多糖 | #8: 多糖 | #9: 多糖 | beta-D-galactopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-2)-[beta-D-galactopyranose- ...beta-D-galactopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-2)-[beta-D-galactopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)]alpha-D-mannopyranose-(1-3)-[beta-D-galactopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-2)-[2-acetamido-2-deoxy-beta-D-glucopyranose-(1-6)]alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta-D-glucopyranose | #10: 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 | #11: 多糖 | #12: 多糖 | 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta- ...2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta-D-glucopyranose | #13: 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 | #14: 多糖 | alpha-D-mannopyranose-(1-6)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1- ...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 | #15: 多糖 | beta-D-galactopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-2)-[beta-D-galactopyranose- ...beta-D-galactopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-2)-[beta-D-galactopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)]alpha-D-mannopyranose-(1-3)-[2-acetamido-2-deoxy-beta-D-glucopyranose-(1-2)-[2-acetamido-2-deoxy-beta-D-glucopyranose-(1-6)]alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta-D-glucopyranose | #16: 糖 | ChemComp-NAG / |
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-詳細
Has protein modification | Y |
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配列の詳細 | V31 WAS FOUND TO HAVE MUTATED TO T31 WHEN THE PLASMID WAS SEQUENCED |
-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: JR-FL ENVDCT IN COMPLEX WITH PGT151 FAB / タイプ: COMPLEX |
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緩衝液 | 名称: 50 MM TRIS, 150 MM NACL, 0.1% DDM, 0.03 MG/ML SODIUM DEOXYCHOLATE pH: 7.4 詳細: 50 MM TRIS, 150 MM NACL, 0.1% DDM, 0.03 MG/ML SODIUM DEOXYCHOLATE |
試料 | 濃度: 3.5 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
試料支持 | 詳細: HOLEY CARBON |
急速凍結 | 装置: HOMEMADE PLUNGER / 凍結剤: ETHANE 詳細: SAMPLES WERE PLUNGED INTO LIQUID ETHANE USING A MANUAL PLUNGER AT RT. |
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電子顕微鏡撮影
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS / 日付: 2015年2月27日 |
電子銃 | 電子線源: ![]() |
電子レンズ | モード: BRIGHT FIELD / 倍率(公称値): 22500 X / 倍率(補正後): 22500 X / 最大 デフォーカス(公称値): 3000 nm / 最小 デフォーカス(公称値): 1200 nm / Cs: 2.7 mm |
試料ホルダ | 傾斜角・最小: 0 ° |
撮影 | 電子線照射量: 32.4 e/Å2 フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) |
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解析
EMソフトウェア |
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CTF補正 | 詳細: WHOLE MICROGRAPH | ||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||
3次元再構成 | 解像度: 4.19 Å / 粒子像の数: 201386 / ピクセルサイズ(公称値): 1.31 Å / ピクセルサイズ(実測値): 1.31 Å 詳細: SUBMISSION BASED ON EXPERIMENTAL DATA FROM EMDB EMD-3308. (DEPOSITION ID: 14201). 対称性のタイプ: POINT | ||||||||||||
原子モデル構築 | プロトコル: OTHER / 空間: REAL / 詳細: REFINEMENT PROTOCOL--EM | ||||||||||||
精密化 | 最高解像度: 4.19 Å | ||||||||||||
精密化ステップ | サイクル: LAST / 最高解像度: 4.2 Å
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