+
データを開く
-
基本情報
登録情報 | ![]() | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
タイトル | Cryo-EM structure of a double loaded human UBA7-UBE2L6-ISG15 thioester mimetic complex (Form 2) | |||||||||
![]() | ||||||||||
![]() |
| |||||||||
![]() | Ligase / Signaling Protein | |||||||||
機能・相同性 | ![]() ISG15 activating enzyme activity / ISG15 transferase activity / 合成酵素; C-S結合を形成; 酸とチオールを結合するもの / positive regulation of protein oligomerization / ISG15-protein conjugation / regulation of type II interferon production / protein localization to mitochondrion / NS1 Mediated Effects on Host Pathways / response to type I interferon / negative regulation of type I interferon-mediated signaling pathway ...ISG15 activating enzyme activity / ISG15 transferase activity / 合成酵素; C-S結合を形成; 酸とチオールを結合するもの / positive regulation of protein oligomerization / ISG15-protein conjugation / regulation of type II interferon production / protein localization to mitochondrion / NS1 Mediated Effects on Host Pathways / response to type I interferon / negative regulation of type I interferon-mediated signaling pathway / E2 ubiquitin-conjugating enzyme / negative regulation of viral genome replication / RSV-host interactions / ubiquitin conjugating enzyme activity / positive regulation of interleukin-10 production / polyubiquitin modification-dependent protein binding / positive regulation of bone mineralization / ubiquitin ligase complex / negative regulation of protein ubiquitination / positive regulation of interferon-beta production / positive regulation of erythrocyte differentiation / ubiquitin binding / Negative regulators of DDX58/IFIH1 signaling / integrin-mediated signaling pathway / Termination of translesion DNA synthesis / DDX58/IFIH1-mediated induction of interferon-alpha/beta / response to virus / protein modification process / PKR-mediated signaling / ISG15 antiviral mechanism / modification-dependent protein catabolic process / protein polyubiquitination / protein tag activity / ubiquitin-protein transferase activity / Interferon alpha/beta signaling / positive regulation of type II interferon production / Antigen processing: Ubiquitination & Proteasome degradation / integrin binding / ubiquitin-dependent protein catabolic process / defense response to virus / protein ubiquitination / defense response to bacterium / Amyloid fiber formation / innate immune response / ubiquitin protein ligase binding / DNA damage response / SARS-CoV-2 activates/modulates innate and adaptive immune responses / extracellular region / nucleoplasm / ATP binding / nucleus / cytoplasm / cytosol 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: 3.2 Å | |||||||||
![]() | Afsar M / Jia L / Ruben EA / Olsen SK | |||||||||
資金援助 | ![]()
| |||||||||
![]() | ![]() タイトル: Cryo-EM structures of Uba7 reveal the molecular basis for ISG15 activation and E1-E2 thioester transfer. 著者: Mohammad Afsar / GuanQun Liu / Lijia Jia / Eliza A Ruben / Digant Nayak / Zuberwasim Sayyad / Priscila Dos Santos Bury / Kristin E Cano / Anindita Nayak / Xiang Ru Zhao / Ankita Shukla / ...著者: Mohammad Afsar / GuanQun Liu / Lijia Jia / Eliza A Ruben / Digant Nayak / Zuberwasim Sayyad / Priscila Dos Santos Bury / Kristin E Cano / Anindita Nayak / Xiang Ru Zhao / Ankita Shukla / Patrick Sung / Elizabeth V Wasmuth / Michaela U Gack / Shaun K Olsen / ![]() 要旨: ISG15 plays a crucial role in the innate immune response and has been well-studied due to its antiviral activity and regulation of signal transduction, apoptosis, and autophagy. ISG15 is a ubiquitin- ...ISG15 plays a crucial role in the innate immune response and has been well-studied due to its antiviral activity and regulation of signal transduction, apoptosis, and autophagy. ISG15 is a ubiquitin-like protein that is activated by an E1 enzyme (Uba7) and transferred to a cognate E2 enzyme (UBE2L6) to form a UBE2L6-ISG15 intermediate that functions with E3 ligases that catalyze conjugation of ISG15 to target proteins. Despite its biological importance, the molecular basis by which Uba7 catalyzes ISG15 activation and transfer to UBE2L6 is unknown as there is no available structure of Uba7. Here, we present cryo-EM structures of human Uba7 in complex with UBE2L6, ISG15 adenylate, and ISG15 thioester intermediate that are poised for catalysis of Uba7-UBE2L6-ISG15 thioester transfer. Our structures reveal a unique overall architecture of the complex compared to structures from the ubiquitin conjugation pathway, particularly with respect to the location of ISG15 thioester intermediate. Our structures also illuminate the molecular basis for Uba7 activities and for its exquisite specificity for ISG15 and UBE2L6. Altogether, our structural, biochemical, and human cell-based data provide significant insights into the functions of Uba7, UBE2L6, and ISG15 in cells. | |||||||||
履歴 |
|
-
構造の表示
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 | ![]() | 166.9 MB | ![]() | |
---|---|---|---|---|
ヘッダ (付随情報) | ![]() ![]() | 18.9 KB 18.9 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 16.5 KB | 表示 | ![]() |
画像 | ![]() | 21.7 KB | ||
マスクデータ | ![]() | 178 MB | ![]() | |
Filedesc metadata | ![]() | 6.5 KB | ||
その他 | ![]() ![]() | 163.2 MB 163.2 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 876.5 KB | 表示 | ![]() |
---|---|---|---|---|
文書・詳細版 | ![]() | 876.1 KB | 表示 | |
XML形式データ | ![]() | 19.3 KB | 表示 | |
CIF形式データ | ![]() | 25.2 KB | 表示 | |
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 | ![]() 8se9MC ![]() 8seaC ![]() 8sebC ![]() 8sv8C ![]() 40410 C: 同じ文献を引用 ( M: このマップから作成された原子モデル |
---|---|
類似構造データ | 類似検索 - 機能・相同性 ![]() |
-
リンク
EMDBのページ | ![]() ![]() |
---|---|
「今月の分子」の関連する項目 |
-
マップ
ファイル | ![]() | ||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
ボクセルのサイズ | X=Y=Z: 0.8332 Å | ||||||||||||||||||||
密度 |
| ||||||||||||||||||||
対称性 | 空間群: 1 | ||||||||||||||||||||
詳細 | EMDB XML:
|
-添付データ
-マスク #1
ファイル | ![]() | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-ハーフマップ: #1
ファイル | emd_40407_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-ハーフマップ: #2
ファイル | emd_40407_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
投影像・断面図 |
| ||||||||||||
密度ヒストグラム |
-
試料の構成要素
-全体 : Cryo-EM structure of a double loaded human UBA7-UBE2L6-ISG15 thio...
全体 | 名称: Cryo-EM structure of a double loaded human UBA7-UBE2L6-ISG15 thioester mimetic complex (Form 2) |
---|---|
要素 |
|
-超分子 #1: Cryo-EM structure of a double loaded human UBA7-UBE2L6-ISG15 thio...
超分子 | 名称: Cryo-EM structure of a double loaded human UBA7-UBE2L6-ISG15 thioester mimetic complex (Form 2) タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#3 |
---|---|
由来(天然) | 生物種: ![]() |
分子量 | 理論値: 160 KDa |
-分子 #1: Ubiquitin-like modifier-activating enzyme 7
分子 | 名称: Ubiquitin-like modifier-activating enzyme 7 / タイプ: protein_or_peptide / ID: 1 / コピー数: 1 / 光学異性体: LEVO |
---|---|
由来(天然) | 生物種: ![]() |
分子量 | 理論値: 111.822102 KDa |
組換発現 | 生物種: ![]() |
配列 | 文字列: MDALDASKLL DEELYSRQLY VLGSPAMQRI QGARVLVSGL QGLGAEVAKN LVLMGVGSLT LHDPHPTCWS DLAAQFLLSE QDLERSRAE ASQELLAQLN RAVQVVVHTG DITEDLLLDF QVVVLTAAKL EEQLKVGTLC HKHGVCFLAA DTRGLVGQLF C DFGEDFTV ...文字列: MDALDASKLL DEELYSRQLY VLGSPAMQRI QGARVLVSGL QGLGAEVAKN LVLMGVGSLT LHDPHPTCWS DLAAQFLLSE QDLERSRAE ASQELLAQLN RAVQVVVHTG DITEDLLLDF QVVVLTAAKL EEQLKVGTLC HKHGVCFLAA DTRGLVGQLF C DFGEDFTV QDPTEAEPLT AAIQHISQGS PGILTLRKGA NTHYFRDGDL VTFSGIEGMV ELNDCDPRSI HVREDGSLEI GD TTTFSRY LRGGAITEVK RPKTVRHKSL DTALLQPHVV AQSSQEVHHA HCLHQAFCAL HKFQHLHGRP PQPWDPVDAE TVV GLARDL EPLKRTEEEP LEEPLDEALV RTVALSSAGV LSPMVAMLGA VAAQEVLKAI SRKFMPLDQW LYFDALDCLP EDGE LLPSP EDCALRGSRY DGQIAVFGAG FQEKLRRQHY LLVGAGAIGC ELLKVFALVG LGAGNSGGLT VVDMDHIERS NLSRQ FLFR SQDVGRPKAE VAAAAARGLN PDLQVIPLTY PLDPTTEHIY GDNFFSRVDG VAAALDSFQA RRYVAARCTH YLKPLL EAG TSGTWGSATV FMPHVTEAYR APASAAASED APYPVCTVRY FPSTAEHTLQ WARHEFEELF RLSAETINHH QQAHTSL AD MDEPQTLTLL KPVLGVLRVR PQNWQDCVAW ALGHWKLCFH YGIKQLLRHF PPNKVLEDGT PFWSGPKQCP QPLEFDTN Q DTHLLYVLAA ANLYAQMHGL PGSQDWTALR ELLKLLPQPD PQQMAPIFAS NLELASASAE FGPEQQKELN KALEVWSVG PPLKPLMFEK DDDSNFHVDF VVAAASLRCQ NYGIPPVNRA QSKRIVGQII PAIATTTAAV AGLLGLELYK VVSGPRPRSA FRHSYLHLA ENYLIRYMPF APAIQTFHHL KWTSWDRLKV PAGQPERTLE SLLAHLQEQH GLRVRILLHG SALLYAAGWS P EKQAQHLP LRVTELVQQL TGQAPAPGQR VLVLELSCEG DDEDTAFPPL HYEL UniProtKB: Ubiquitin-like modifier-activating enzyme 7 |
-分子 #2: Ubiquitin-like protein ISG15
分子 | 名称: Ubiquitin-like protein ISG15 / タイプ: protein_or_peptide / ID: 2 / コピー数: 2 / 光学異性体: LEVO |
---|---|
由来(天然) | 生物種: ![]() |
分子量 | 理論値: 17.163701 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MGWDLTVKML AGNEFQVSLS SSMSVSELKA QITQKIGVHA FQQRLAVHPS GVALQDRVPL ASQGLGPGST VLLVVDKCDE PLSILVRNN KGRSSTYEVR LTQTVAHLKQ QVSGLEGVQD DLFWLTFEGK PLEDQLPLGE YGLKPLSTVF MNLRLRGG UniProtKB: Ubiquitin-like protein ISG15 |
-分子 #3: Ubiquitin/ISG15-conjugating enzyme E2 L6
分子 | 名称: Ubiquitin/ISG15-conjugating enzyme E2 L6 / タイプ: protein_or_peptide / ID: 3 / コピー数: 1 / 光学異性体: LEVO / EC番号: E2 ubiquitin-conjugating enzyme |
---|---|
由来(天然) | 生物種: ![]() |
分子量 | 理論値: 17.643377 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MASMRVVKEL EDLQKKPPPY LRNLSSDDAN VLVWHALLLP DQPPYHLKAF NLRISFPPEY PFKPPMIKFT TKIYHPNVDE NGQICLPII SSENWKPSTK TSQVLEALNV LVNRPNIREP KRMDLADLLT QNPELFRKNA EEFTLRFGVD RPS UniProtKB: Ubiquitin/ISG15-conjugating enzyme E2 L6 |
-分子 #4: ADENOSINE MONOPHOSPHATE
分子 | 名称: ADENOSINE MONOPHOSPHATE / タイプ: ligand / ID: 4 / コピー数: 1 / 式: AMP |
---|---|
分子量 | 理論値: 347.221 Da |
Chemical component information | ![]() ChemComp-AMP: |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
---|---|
![]() | 単粒子再構成法 |
試料の集合状態 | particle |
-
試料調製
緩衝液 | pH: 7.5 |
---|---|
凍結 | 凍結剤: ETHANE |
-
電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
---|---|
撮影 | フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 平均電子線量: 80.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
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): 2.888 µm / 最小 デフォーカス(公称値): 0.1 µm |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |