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基本情報
登録情報 | ![]() | ||||||||||||||||||
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タイトル | Coagulation factor VIII bound to a patient-derived anti-C1 domain antibody inhibitor | ||||||||||||||||||
![]() | Blood coagulation factor VIII bound to patient-derived anti-C1 domain antibody inhibitor | ||||||||||||||||||
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![]() | Immune system / BLOOD CLOTTING | ||||||||||||||||||
機能・相同性 | ![]() Defective F8 accelerates dissociation of the A2 domain / Defective F8 binding to the cell membrane / Defective F8 secretion / Defective F8 sulfation at Y1699 / Gamma carboxylation, hypusinylation, hydroxylation, and arylsulfatase activation / Defective F8 binding to von Willebrand factor / blood coagulation, intrinsic pathway / Cargo concentration in the ER / Defective factor IX causes thrombophilia / Defective cofactor function of FVIIIa variant ...Defective F8 accelerates dissociation of the A2 domain / Defective F8 binding to the cell membrane / Defective F8 secretion / Defective F8 sulfation at Y1699 / Gamma carboxylation, hypusinylation, hydroxylation, and arylsulfatase activation / Defective F8 binding to von Willebrand factor / blood coagulation, intrinsic pathway / Cargo concentration in the ER / Defective factor IX causes thrombophilia / Defective cofactor function of FVIIIa variant / Defective F9 variant does not activate FX / COPII-mediated vesicle transport / COPII-coated ER to Golgi transport vesicle / Defective F8 cleavage by thrombin / Common Pathway of Fibrin Clot Formation / Intrinsic Pathway of Fibrin Clot Formation / endoplasmic reticulum-Golgi intermediate compartment membrane / platelet alpha granule lumen / acute-phase response / Golgi lumen / blood coagulation / Platelet degranulation / oxidoreductase activity / copper ion binding / endoplasmic reticulum lumen / extracellular space / extracellular region / plasma membrane 類似検索 - 分子機能 | ||||||||||||||||||
生物種 | ![]() | ||||||||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.23 Å | ||||||||||||||||||
![]() | Childers KC / Davulcu O / Haynes RM / Lollar P / Doering CB / Coxon CH / Spiegel PC | ||||||||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Structure of coagulation factor VIII bound to a patient-derived anti-C1 domain antibody inhibitor. 著者: Kenneth C Childers / Nathan G Avery / Kevin A Estrada Alamo / Omar Davulcu / Rose Marie Haynes / Pete Lollar / Christopher B Doering / Carmen H Coxon / P Clint Spiegel / ![]() ![]() 要旨: The development of pathogenic antibody inhibitors against coagulation factor VIII (FVIII) occurs in ∼30% of patients with congenital hemophilia A receiving FVIII replacement therapy, as well as in ...The development of pathogenic antibody inhibitors against coagulation factor VIII (FVIII) occurs in ∼30% of patients with congenital hemophilia A receiving FVIII replacement therapy, as well as in all cases of acquired hemophilia A. KM33 is an anti-C1 domain antibody inhibitor previously isolated from a patient with severe hemophilia A. In addition to potently blocking FVIII binding to von Willebrand factor and phospholipid surfaces, KM33 disrupts FVIII binding to lipoprotein receptor-related protein 1 (LRP1), which drives FVIII hepatic clearance and antigen presentation in dendritic cells. Here, we report on the structure of FVIII bound to NB33, a recombinant derivative of KM33, via single-particle cryo-electron microscopy. Structural analysis revealed that the NB33 epitope localizes to the FVIII residues R2090-S2094 and I2158-R2159, which constitute membrane-binding loops in the C1 domain. Further analysis revealed that multiple FVIII lysine and arginine residues, previously shown to mediate binding to LRP1, dock onto an acidic cleft at the NB33 variable domain interface, thus blocking a putative LRP1 binding site. Together, these results demonstrate a novel mechanism of FVIII inhibition by a patient-derived antibody inhibitor and provide structural evidence for engineering FVIII with reduced LRP1-mediated clearance. | ||||||||||||||||||
履歴 |
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構造の表示
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マップデータ | ![]() | 167.7 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 21.9 KB 21.9 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 12 KB | 表示 | ![]() |
画像 | ![]() | 102.8 KB | ||
その他 | ![]() ![]() | 165.1 MB 165.1 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 8g6iMC M: このマップから作成された原子モデル 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 ![]() |
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リンク
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||
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注釈 | Blood coagulation factor VIII bound to patient-derived anti-C1 domain antibody inhibitor | ||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 0.788 Å | ||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-ハーフマップ: Blood coagulation factor VIII bound to patient-derived anti-C1...
ファイル | emd_29770_half_map_1.map | ||||||||||||
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注釈 | Blood coagulation factor VIII bound to patient-derived anti-C1 domain antibody inhibitor; half-map A | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: Blood coagulation factor VIII bound to patient-derived anti-C1...
ファイル | emd_29770_half_map_2.map | ||||||||||||
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注釈 | Blood coagulation factor VIII bound to patient-derived anti-C1 domain antibody inhibitor; half-map B | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
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試料の構成要素
-全体 : Structure of coagulation factor VIII bound to a patient-derived a...
全体 | 名称: Structure of coagulation factor VIII bound to a patient-derived anti-C1 domain antibody inhibitor |
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要素 |
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-超分子 #1: Structure of coagulation factor VIII bound to a patient-derived a...
超分子 | 名称: Structure of coagulation factor VIII bound to a patient-derived anti-C1 domain antibody inhibitor タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: all 詳細: Coagulation factor VIII (molecule 1) is a chimera of human+pig proteins which was expressed in CHO cells. |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 220 KDa |
-分子 #1: Coagulation factor VIII chimera from human and pig
分子 | 名称: Coagulation factor VIII chimera from human and pig / タイプ: protein_or_peptide / ID: 1 詳細: This protein is a chimera of human+pig coagulation factor VIII (PDB: 6MF0). コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 168.287047 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MQLELSTCVF LCLLPLGFSA IRRYYLGAVE LSWDYRQSEL LRELHVDTRF PATAPGALPL GPSVLYKKTV FVEFTDQLFS VARPRPPWM GLLGPTIQAE VYDTVVVTLK NMASHPVSLH AVGVSFWKSS EGAEYEDHTS QREKEDDKVL PGKSQTYVWQ V LKENGPTA ...文字列: MQLELSTCVF LCLLPLGFSA IRRYYLGAVE LSWDYRQSEL LRELHVDTRF PATAPGALPL GPSVLYKKTV FVEFTDQLFS VARPRPPWM GLLGPTIQAE VYDTVVVTLK NMASHPVSLH AVGVSFWKSS EGAEYEDHTS QREKEDDKVL PGKSQTYVWQ V LKENGPTA SDPPCLTYSY LSHVDLVKDL NSGLIGALLV CREGSLTRER TQNLHEFVLL FAVFDEGKSW HSARNDSWTR AM DPAPARA QPAMHTVNGY VNRSLPGLIG CHKKSVYWHV IGMGTSPEVH SIFLEGHTFL VRHHRQASLE ISPLTFLTAQ TFL MDLGQF LLFCHISSHH HGGMEAHVRV ESCAEEPQLR RKADEEEDYD DNLYDSDMDV VRLDGDDVSP FIQIRSVAKK HPKT WVHYI AAEEEDWDYA PLVLAPDDRS YKSQYLNNGP QRIGRKYKKV RFMAYTDETF KTREAIQHES GILGPLLYGE VGDTL LIIF KNQASRPYNI YPHGITDVRP LYSRRLPKGV KHLKDFPILP GEIFKYKWTV TVEDGPTKSD PRCLTRYYSS FVNMER DLA SGLIGPLLIC YKESVDQRGN QIMSDKRNVI LFSVFDENRS WYLTENIQRF LPNPAGVQLE DPEFQASNIM HSINGYV FD SLQLSVCLHE VAYWYILSIG AQTDFLSVFF SGYTFKHKMV YEDTLTLFPF SGETVFMSME NPGLWILGCH NSDFRNRG M TALLKVSSCD KNTGDYYEDS YEDISAYLLS KNNAIEPRSF AQNSRPPSAS APKPPVLRRH QRDISLPTFQ PEEDKMDYD DIFSTETKGE DFDIYGEDEN QDPRSFQKRT RHYFIAAVEQ LWDYGMSESP RALRNRAQNG EVPRFKKVVF REFADGSFTQ PSYRGELNK HLGLLGPYIR AEVEDNIMVT FKNQASRPYS FYSSLISYPD DQEQGAEPRH NFVQPNETRT YFWKVQHHMA P TEDEFDCK AWAYFSDVDL EKDVHSGLIG PLLICRANTL NAAHGRQVTV QEFALFFTIF DETKSWYFTE NVERNCRAPC HL QMEDPTL KENYRFHAIN GYVMDTLPGL VMAQNQRIRW YLLSMGSNEN IHSIHFSGHV FSVRKKEEYK MAVYNLYPGV FET VEMLPS KVGIWRIECL IGEHLQAGMS TTFLVYSKKC QTPLGMASGH IRDFQITASG QYGQWAPKLA RLHYSGSINA WSTK EPFSW IKVDLLAPMI IHGIKTQGAR QKFSSLYISQ FIIMYSLDGK KWQTYRGNST GTLMVFFGNV DSSGIKHNIF NPPII ARYI RLHPTHYSIR STLRMELMGC DLNSCSMPLG MESKAISDAQ ITASSYFTNM FATWSPSKAR LHLQGRSNAW RPQVNN PKE WLQVDFQKTM KVTGVTTQGV KSLLTSMYVK EFLISSSQDG HQWTLFFQNG KVKVFQGNQD SFTPVVNSLD PPLLTRY LR IHPQSWVHQI ALRMEVLGCE AQDLY UniProtKB: Coagulation factor VIII, Coagulation factor VIII, Coagulation factor VIII, Coagulation factor VIII |
-分子 #2: NB33 light chain
分子 | 名称: NB33 light chain / 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: ![]() |
分子量 | 理論値: 22.972436 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: DIQMTQSPSS LSASVGDTVT IACRASRDIR NDLAWYQQKP GKAPKLLIYA TSRLQSGVPS RFSGSGSFTD FTLTINSLQP DDSATYYCL QDSDYPLTFG GGTKVDIKGT VAAPSVFIFP PSDEQLKSGT ASVVCLLNNF YPREAKVQWK VDNALQSGNS Q ESVTEQDS ...文字列: DIQMTQSPSS LSASVGDTVT IACRASRDIR NDLAWYQQKP GKAPKLLIYA TSRLQSGVPS RFSGSGSFTD FTLTINSLQP DDSATYYCL QDSDYPLTFG GGTKVDIKGT VAAPSVFIFP PSDEQLKSGT ASVVCLLNNF YPREAKVQWK VDNALQSGNS Q ESVTEQDS KDSTYSLSST LTLSKADYEK HKVYACEVTH QGLSSPVTKS FNR |
-分子 #3: NB33 heavy chain
分子 | 名称: NB33 heavy chain / タイプ: protein_or_peptide / ID: 3 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 22.94566 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: EVQLVESGGG VVQPGRSLRL SCVDSGLTFS SYGMHWVRQA PGAGLEWVAV ISYDGNDKYY ADSVKGRFAI SRDNAKNTLY LQMNSLTIE DTAVYYCAKD LIESNIAEAF WGQGTLVTVS SKGPSVFPLA PCSRSTSEST AALGCLVKDY FPEPVTVSWN S GALTSGVH ...文字列: EVQLVESGGG VVQPGRSLRL SCVDSGLTFS SYGMHWVRQA PGAGLEWVAV ISYDGNDKYY ADSVKGRFAI SRDNAKNTLY LQMNSLTIE DTAVYYCAKD LIESNIAEAF WGQGTLVTVS SKGPSVFPLA PCSRSTSEST AALGCLVKDY FPEPVTVSWN S GALTSGVH TFPAVLQSSG LYSLSSVVTV PSSSLGTATY TCNVDHKPSN TKVDKRV |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 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 | 0.1 mg/mL | |||||||||
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緩衝液 | pH: 7.4 構成要素:
詳細: 150 mM NaCl, 20 mM HEPES pH 7.4 | |||||||||
グリッド | モデル: EMS Lacey Carbon / メッシュ: 300 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: LACEY / 前処理 - タイプ: GLOW DISCHARGE | |||||||||
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 I | |||||||||
詳細 | This sample was monodisperse. |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 撮影したグリッド数: 1 / 実像数: 6993 / 平均露光時間: 1.565 sec. / 平均電子線量: 50.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | C2レンズ絞り径: 100.0 µm / 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): 2.0 µm / 最小 デフォーカス(公称値): 0.8 µm |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |