+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-25581 | |||||||||
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タイトル | cryoEM reconstruction of the HIV gp140 in complex with the extracellular domains of CD4 and the adnectin domain of Combinectin. The gp140 and CD4 coordinates from entry 6EDU were rigid body fitted to the EM map along withe the crystal structure of CD4+adnectin | |||||||||
マップデータ | ||||||||||
試料 |
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機能・相同性 | 機能・相同性情報 helper T cell enhancement of adaptive immune response / interleukin-16 binding / interleukin-16 receptor activity / maintenance of protein location in cell / T cell selection / MHC class II protein binding / cellular response to granulocyte macrophage colony-stimulating factor stimulus / interleukin-15-mediated signaling pathway / positive regulation of monocyte differentiation / Nef Mediated CD4 Down-regulation ...helper T cell enhancement of adaptive immune response / interleukin-16 binding / interleukin-16 receptor activity / maintenance of protein location in cell / T cell selection / MHC class II protein binding / cellular response to granulocyte macrophage colony-stimulating factor stimulus / interleukin-15-mediated signaling pathway / positive regulation of monocyte differentiation / Nef Mediated CD4 Down-regulation / Alpha-defensins / positive regulation of kinase activity / regulation of T cell activation / extracellular matrix structural constituent / T cell receptor complex / Other interleukin signaling / enzyme-linked receptor protein signaling pathway / Translocation of ZAP-70 to Immunological synapse / Phosphorylation of CD3 and TCR zeta chains / regulation of calcium ion transport / macrophage differentiation / T cell differentiation / Generation of second messenger molecules / PD-1 signaling / positive regulation of protein kinase activity / Binding and entry of HIV virion / coreceptor activity / cell surface receptor protein tyrosine kinase signaling pathway / positive regulation of interleukin-2 production / positive regulation of calcium-mediated signaling / T cell activation / protein tyrosine kinase binding / Vpu mediated degradation of CD4 / calcium-mediated signaling / clathrin-coated endocytic vesicle membrane / positive regulation of peptidyl-tyrosine phosphorylation / transmembrane signaling receptor activity / positive regulation of T cell activation / Cargo recognition for clathrin-mediated endocytosis / Downstream TCR signaling / MHC class II protein complex binding / Clathrin-mediated endocytosis / signaling receptor activity / virus receptor activity / defense response to Gram-negative bacterium / positive regulation of canonical NF-kappaB signal transduction / adaptive immune response / positive regulation of MAPK cascade / positive regulation of ERK1 and ERK2 cascade / early endosome / positive regulation of viral entry into host cell / cell surface receptor signaling pathway / cell adhesion / immune response / positive regulation of protein phosphorylation / membrane raft / endoplasmic reticulum lumen / external side of plasma membrane / lipid binding / endoplasmic reticulum membrane / protein kinase binding / positive regulation of DNA-templated transcription / enzyme binding / signal transduction / protein homodimerization activity / zinc ion binding / identical protein binding / plasma membrane 類似検索 - 分子機能 | |||||||||
生物種 | HIV-1 06TG.HT008 (ヒト免疫不全ウイルス) / Homo sapiens (ヒト) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 8.7 Å | |||||||||
データ登録者 | Concha NO / William SP / Wenzel DL | |||||||||
資金援助 | 米国, 1件
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引用 | ジャーナル: J Mol Biol / 年: 2022 タイトル: Novel Bent Conformation of CD4 Induced by HIV-1 Inhibitor Indirectly Prevents Productive Viral Attachment. 著者: David Wensel / Shawn Williams / David P Dixon / Paris Ward / Patti McCormick / Nestor Concha / Eugene Stewart / Xuan Hong / Charles Mazzucco / Shreya Pal / Bo Ding / Christoph Fellinger / Mark Krystal / 要旨: GSK3732394 is a multi-specific biologic inhibitor of HIV entry currently under clinical evaluation. A key component of this molecule is an adnectin (6940_B01) that binds to CD4 and inhibits ...GSK3732394 is a multi-specific biologic inhibitor of HIV entry currently under clinical evaluation. A key component of this molecule is an adnectin (6940_B01) that binds to CD4 and inhibits downstream actions of gp160. Studies were performed to determine the binding site of the adnectin on CD4 and to understand the mechanism of inhibition. Using hydrogen-deuterium exchange with mass spectrometry (HDX), CD4 peptides showed differential rates of deuteration (either enhanced or slowed) in the presence of the adnectin that mapped predominantly to the interface of domains 2 and 3 (D2-D3). In addition, an X-ray crystal structure of an ibalizumab Fab/CD4(D1-D4)/adnectin complex revealed an extensive interface between the adnectin and residues on CD4 domains D2-D4 that stabilize a novel T-shaped CD4 conformation. A cryo-EM map of the gp140/CD4/GSK3732394 complex clearly shows the bent conformation for CD4 while bound to gp140. Mutagenic analyses on CD4 confirmed that amino acid F202 forms a key interaction with the adnectin. In addition, amino acid L151 was shown to be a critical indirect determinant of the specificity for binding to the human CD4 protein over related primate CD4 molecules, as it appears to modulate CD4's flexibility to adopt the adnectin-bound conformation. The significant conformational change of CD4 upon adnectin binding brings the D1 domain of CD4 in proximity to the host cell membrane surface, thereby re-orienting the gp120 binding site in a direction that is inaccessible to incoming virus due to a steric clash between gp160 trimers on the virus surface and the target cell membrane. | |||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_25581.map.gz | 1.6 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-25581-v30.xml emd-25581.xml | 21.7 KB 21.7 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_25581_fsc.xml | 6.5 KB | 表示 | FSCデータファイル |
画像 | emd_25581.png | 65 KB | ||
マスクデータ | emd_25581_msk_1.map | 22.2 MB | マスクマップ | |
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-25581 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-25581 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_25581_validation.pdf.gz | 373.8 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_25581_full_validation.pdf.gz | 373.4 KB | 表示 | |
XML形式データ | emd_25581_validation.xml.gz | 8.3 KB | 表示 | |
CIF形式データ | emd_25581_validation.cif.gz | 10.7 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25581 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25581 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_25581.map.gz / 形式: CCP4 / 大きさ: 1.8 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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投影像・断面図 | 画像のコントロール
画像は Spider により作成 これらの図は立方格子座標系で作成されたものです | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 2.4624 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
-マスク #1
ファイル | emd_25581_msk_1.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
-全体 : Ternary complex of the HIV-1 gp140 trimer in complex with the ext...
全体 | 名称: Ternary complex of the HIV-1 gp140 trimer in complex with the extracellular domains (domains 1-4) of CD4 and an adnectin domain of Combinectin |
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要素 |
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-超分子 #1: Ternary complex of the HIV-1 gp140 trimer in complex with the ext...
超分子 | 名称: Ternary complex of the HIV-1 gp140 trimer in complex with the extracellular domains (domains 1-4) of CD4 and an adnectin domain of Combinectin タイプ: complex / キメラ: Yes / ID: 1 / 親要素: 0 / 含まれる分子: all 詳細: the gp140 trimer at 0.67 mg/ml was mixed with soluble CD4 first, and then with Combinection in a 1.25:1 ratio. The sample was incubated overnight at 4 degC. The sample was concentrated to ...詳細: the gp140 trimer at 0.67 mg/ml was mixed with soluble CD4 first, and then with Combinection in a 1.25:1 ratio. The sample was incubated overnight at 4 degC. The sample was concentrated to 35ul before injecting into a Phenomenex 4.6 x 300 mm 3um bead 500Angstroms pore Yara SEC column equilibrated with 20mM Hepes pH7.5, 200 mM KCl, 5mM MgCl2. The complexes were eluted with a flow rate of 0.2 ml/min and fractions collected every 20 seconds into a Thermo-Fisher UV transparent 96-well plates. The absorbance at 280nm was measured for each fraction using a Spectramax plate scanner. The high-molecular weight complex appeared to be comprised of gp140:CD4:Combinectin in a 1:1:1 ratio. The sample used for cryo-EM studies was the single peak fraction of gp140/CD4/Combinectin (0.076 mg/ml) |
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-分子 #1: BG505 SOSIP.664 gp140
分子 | 名称: BG505 SOSIP.664 gp140 / タイプ: protein_or_peptide / ID: 1 / コピー数: 3 / 光学異性体: LEVO |
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由来(天然) | 生物種: HIV-1 06TG.HT008 (ヒト免疫不全ウイルス) |
分子量 | 理論値: 72.830398 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: AENLWVTVYY GVPVWKDAET TLFCASDAKA YETEKHNVWA THACVPTDPN PQEIHLENVT EEFNMWKNNM VEQMHTDIIS LWDQSLKPC VKLTPLCVTL NCTNVTNNIT DDMRGELKNC SFNMTTELRD KKQKVYSLFY RLDVVQINEN QGNRSNNSNK E YRLINCNT ...文字列: AENLWVTVYY GVPVWKDAET TLFCASDAKA YETEKHNVWA THACVPTDPN PQEIHLENVT EEFNMWKNNM VEQMHTDIIS LWDQSLKPC VKLTPLCVTL NCTNVTNNIT DDMRGELKNC SFNMTTELRD KKQKVYSLFY RLDVVQINEN QGNRSNNSNK E YRLINCNT SAITQACPKV SFEPIPIHYC APAGFAILKC KDKKFNGTGP CPSVSTVQCT HGIKPVVSTQ LLLNGSLAEE EV MIRSENI TNNAKNILVQ FNTPVQINCT RPNNNTRKSI RIGPGQAFYA TGDIIGDIRQ AHCNVSKATW NETLGKVVKQ LRK HFGNNT IIRFANSSGG DLEVTTHSFN CGGEFFYCNT SGLFNSTWIS NTSVQGSNST GSNDSITLPC RIKQIINMWQ RIGQ AMYAP PIQGVIRCVS NITGLILTRD GGSTNSTTET FRPGGGDMRD NWRSELYKYK VVKIEPLGVA PTRCKRRVVG RRRRR RAVG IGAVFLGFLG AAGSTMGAAS MTLTVQARNL LSGIVQQQSN LLRAPEAQQH LLKLTVWGIK QLQARVLAVE RYLRDQ QLL GIWGCSGKLI CCTNVPWNSS WSNRNLSEIW DNMTWLQWDK EISNYTQIIY GLLEESQNQQ EKNEQDLLAL DGSGSGG SG HHHHHHHH |
-分子 #2: BG505 SOSIP.664 gp140
分子 | 名称: BG505 SOSIP.664 gp140 / タイプ: protein_or_peptide / ID: 2 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: HIV-1 06TG.HT008 (ヒト免疫不全ウイルス) |
分子量 | 理論値: 3.337105 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK) |
-分子 #3: BG505 SOSIP.664 gp140
分子 | 名称: BG505 SOSIP.664 gp140 / タイプ: protein_or_peptide / ID: 3 / コピー数: 2 / 光学異性体: LEVO |
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由来(天然) | 生物種: HIV-1 06TG.HT008 (ヒト免疫不全ウイルス) |
分子量 | 理論値: 3.252 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) |
-分子 #4: T-cell surface glycoprotein CD4
分子 | 名称: T-cell surface glycoprotein CD4 / タイプ: protein_or_peptide / ID: 4 / コピー数: 3 / 光学異性体: LEVO |
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由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 41.385449 KDa |
組換発現 | 生物種: Spodoptera frugiperda (ツマジロクサヨトウ) |
配列 | 文字列: KKVVLGKKGD TVELTCTASQ KKSIQFHWKN SNQIKILGNQ GSFLTKGPSK LNDRADSRRS LWDQGNFPLI IKNLKIEDSD TYICEVEDQ KEEVQLLVFG LTANSDTHLL QGQSLTLTLE SPPGSSPSVQ CRSPRGKNIQ GGKTLSVSQL ELQDSGTWTC T VLQNQKKV ...文字列: KKVVLGKKGD TVELTCTASQ KKSIQFHWKN SNQIKILGNQ GSFLTKGPSK LNDRADSRRS LWDQGNFPLI IKNLKIEDSD TYICEVEDQ KEEVQLLVFG LTANSDTHLL QGQSLTLTLE SPPGSSPSVQ CRSPRGKNIQ GGKTLSVSQL ELQDSGTWTC T VLQNQKKV EFKIDIVVLA FQKASSIVYK KEGEQVEFSF PLAFTVEKLT GSGELWWQAE RASSSKSWIT FDLKNKEVSV KR VTQDPKL QMGKKLPLHL TLPQALPQYA GSGNLTLALE AKTGKLHQEV NLVVMRATQL QKNLTCEVWG PTSPKLMLSL KLE NKEAKV SKREKAVWVL NPEAGMWQCL LSDSGQVLLE SNIKVLPTHH HHHH |
-分子 #5: Adnectin
分子 | 名称: Adnectin / タイプ: protein_or_peptide / ID: 5 詳細: The native 10th fibronectin type III (10Fn3) domain from human fibronectin was used as the template for construction of novel libraries of Adnectin variants (Wenzel, D., et al. (2017) ...詳細: The native 10th fibronectin type III (10Fn3) domain from human fibronectin was used as the template for construction of novel libraries of Adnectin variants (Wenzel, D., et al. (2017) Antimicrobial Agents and Chemotherapy, Vol 61(8), e00508-17, pp 1-19) コピー数: 3 / 光学異性体: LEVO |
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由来(天然) | 生物種: Homo sapiens (ヒト) |
分子量 | 理論値: 11.959172 KDa |
組換発現 | 生物種: Escherichia coli (大腸菌) |
配列 | 文字列: GVSDVPRDLE VVAATPTSLL ISWDAPAVTV HSYHIQYWPL GSYQRYQVFS VPGSKSTATI SGLKPGVEYQ IRVYAETGGA DSDQSMGWI QIGYRTEGDK PSQHHHHHH |
-分子 #6: BG505 SOSIP.664 gp140
分子 | 名称: BG505 SOSIP.664 gp140 / タイプ: protein_or_peptide / ID: 6 / コピー数: 3 / 光学異性体: LEVO |
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由来(天然) | 生物種: HIV-1 06TG.HT008 (ヒト免疫不全ウイルス) |
分子量 | 理論値: 2.996685 KDa |
組換発現 | 生物種: Homo sapiens (ヒト) |
配列 | 文字列: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK) |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
濃度 | 0.08 mg/mL | ||||||||
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緩衝液 | pH: 7.5 / 構成要素:
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グリッド | モデル: Quantifoil R1.2/1.3 / 材質: COPPER / メッシュ: 400 / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 雰囲気: AIR / 前処理 - 気圧: 30.0 kPa 詳細: glow discharged for 20 sec using a EasyGlo (20mA, 0.3 mBar) | ||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 K / 装置: FEI VITROBOT MARK IV | ||||||||
詳細 | the gp140/CD4/Combinectin (0.076 mg/ml), 3.5uL were applied to Quantifoil 1.2/1.3 Cu 400 mesh grids that had previously been glow discharged for 20 sec using a EasyGlo (20mA, 0.3 mBar). The grids were plunged into liquid ethane using a Vitrobot plunger (chamber set to 4 degC and 80% RH) immediately after blotting from 2.5 to 4 sec |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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特殊光学系 | 位相板: OTHER / 球面収差補正装置: Cs corrector |
撮影 | フィルム・検出器のモデル: FEI FALCON III (4k x 4k) 検出モード: COUNTING / デジタル化 - サイズ - 横: 4096 pixel / デジタル化 - サイズ - 縦: 4096 pixel / デジタル化 - サンプリング間隔: 11.0 µm / 実像数: 1078 / 平均露光時間: 75.7 sec. / 平均電子線量: 1.022 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 3.4 µm 最小 デフォーカス(公称値): 1.4000000000000001 µm |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
+画像解析
-原子モデル構築 1
精密化 | プロトコル: RIGID BODY FIT |
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得られたモデル | PDB-7t0o: |