+データを開く
-基本情報
登録情報 | データベース: SASBDB / ID: SASDEK3 |
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試料 | The neutrophil cytosol factor 1 (p47phox) subunit of phagocyte NADPH oxidase
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機能・相同性 | 機能・相同性情報 regulation of respiratory burst involved in inflammatory response / superoxide-generating NADPH oxidase activator activity / reactive oxygen species biosynthetic process / positive regulation of epidermal growth factor-activated receptor activity / phagolysosome / superoxide-generating NAD(P)H oxidase activity / Cross-presentation of particulate exogenous antigens (phagosomes) / NADPH oxidase complex / respiratory burst / cellular response to testosterone stimulus ...regulation of respiratory burst involved in inflammatory response / superoxide-generating NADPH oxidase activator activity / reactive oxygen species biosynthetic process / positive regulation of epidermal growth factor-activated receptor activity / phagolysosome / superoxide-generating NAD(P)H oxidase activity / Cross-presentation of particulate exogenous antigens (phagosomes) / NADPH oxidase complex / respiratory burst / cellular response to testosterone stimulus / ROS and RNS production in phagocytes / phosphatidylinositol-3,4-bisphosphate binding / superoxide anion generation / protein targeting to membrane / positive regulation of p38MAPK cascade / superoxide metabolic process / Detoxification of Reactive Oxygen Species / RHO GTPases Activate NADPH Oxidases / RAC2 GTPase cycle / RAC3 GTPase cycle / cellular defense response / RAC1 GTPase cycle / cellular response to cadmium ion / phosphatidylinositol binding / cellular response to glucose stimulus / positive regulation of JNK cascade / VEGFA-VEGFR2 Pathway / SH3 domain binding / cytoplasmic side of plasma membrane / cellular response to reactive oxygen species / electron transfer activity / positive regulation of phosphatidylinositol 3-kinase/protein kinase B signal transduction / innate immune response / neuronal cell body / dendrite / positive regulation of DNA-templated transcription / membrane / plasma membrane / cytosol / cytoplasm 類似検索 - 分子機能 |
生物種 | Homo sapiens (ヒト) |
引用 | ジャーナル: J Biol Chem / 年: 2019 タイトル: Quantitative live-cell imaging and 3D modeling reveal critical functional features in the cytosolic complex of phagocyte NADPH oxidase. 著者: Cornelia S Ziegler / Leïla Bouchab / Marc Tramier / Dominique Durand / Franck Fieschi / Sophie Dupré-Crochet / Fabienne Mérola / Oliver Nüße / Marie Erard / 要旨: Phagocyte NADPH oxidase produces superoxide anions, a precursor of reactive oxygen species (ROS) critical for host responses to microbial infections. However, uncontrolled ROS production contributes ...Phagocyte NADPH oxidase produces superoxide anions, a precursor of reactive oxygen species (ROS) critical for host responses to microbial infections. However, uncontrolled ROS production contributes to inflammation, making NADPH oxidase a major drug target. It consists of two membranous (Nox2 and p22) and three cytosolic subunits (p40, p47, and p67) that undergo structural changes during enzyme activation. Unraveling the interactions between these subunits and the resulting conformation of the complex could shed light on NADPH oxidase regulation and help identify inhibition sites. However, the structures and the interactions of flexible proteins comprising several well-structured domains connected by intrinsically disordered protein segments are difficult to investigate by conventional techniques such as X-ray crystallography, NMR, or cryo-EM. Here, we developed an analytical strategy based on FRET-fluorescence lifetime imaging (FLIM) and fluorescence cross-correlation spectroscopy (FCCS) to structurally and quantitatively characterize NADPH oxidase in live cells. We characterized the inter- and intramolecular interactions of its cytosolic subunits by elucidating their conformation, stoichiometry, interacting fraction, and affinities in live cells. Our results revealed that the three subunits have a 1:1:1 stoichiometry and that nearly 100% of them are present in complexes in living cells. Furthermore, combining FRET data with small-angle X-ray scattering (SAXS) models and published crystal structures of isolated domains and subunits, we built a 3D model of the entire cytosolic complex. The model disclosed an elongated complex containing a flexible hinge separating two domains ideally positioned at one end of the complex and critical for oxidase activation and interactions with membrane components. |
登録者 |
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-構造の表示
構造ビューア | 分子: MolmilJmol/JSmol |
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-ダウンロードとリンク
-モデル
モデル #2309 | タイプ: atomic / カイ2乗値: 1.245 Omokage検索でこの集合体の類似形状データを探す (詳細) |
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-試料
試料 | 名称: The neutrophil cytosol factor 1 (p47phox) subunit of phagocyte NADPH oxidase 試料濃度: 1.00-4.00 |
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バッファ | 名称: 50 mM HEPES, 100 mM NaCl, 1 mM EDTA, 2 mM DTT, 5% glycerol pH: 7.5 |
要素 #1262 | 名称: p47phox / タイプ: protein / 記述: Neutrophil cytosol factor 1 / 分子量: 46.364 / 分子数: 1 / 由来: Homo sapiens / 参照: UniProt: P14598 配列: GPLGSPNSAR MGDTFIRHIA LLGFEKRFVP SQHYVYMFLV KWQDLSEKVV YRRFTEIYEF HKTLKEMFPI EAGAINPENR IIPHLPAPKW FDGQRAAENR QGTLTEYCST LMSLPTKISR CPHLLDFFKV RPDDLKLPTD NQTKKPETYL MPKDGKSTAT DITGPIILQT ...配列: GPLGSPNSAR MGDTFIRHIA LLGFEKRFVP SQHYVYMFLV KWQDLSEKVV YRRFTEIYEF HKTLKEMFPI EAGAINPENR IIPHLPAPKW FDGQRAAENR QGTLTEYCST LMSLPTKISR CPHLLDFFKV RPDDLKLPTD NQTKKPETYL MPKDGKSTAT DITGPIILQT YRAIADYEKT SGSEMALSTG DVVEVVEKSE SGWWFCQMKA KRGWIPASFL EPLDSPDETE DPEPNYAGEP YVAIKAYTAV EGDEVSLLEG EAVEVIHKLL DGWWVIRKDD VTGYFPSMYL QKSGQDVSQA QRQIKRGAPP RRSSIRNAHS IHQRSRKRLS QDAYRRNSVR FLQQRRRQAR PGPQSPGSPL EEERQTQRSK PQPAVPPRPS ADLILNRCSE STKRKLASAV CGRIVTD |
-実験情報
ビーム | 設備名称: SOLEIL SWING / 地域: Saint-Aubin / 国: France / 線源: X-ray synchrotron / 波長: 0.1 Å / スペクトロメータ・検出器間距離: 1.783 mm | ||||||||||||||||||||||||||||||||||||||||||
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検出器 | 名称: AVIEX PCCD170170 / タイプ: CCD | ||||||||||||||||||||||||||||||||||||||||||
スキャン | タイトル: Neutrophil cytosol factor 1 (p47phox) subunit of the phagocyte NADPH oxidase 測定日: 2008年4月23日 / 保管温度: 10 °C / セル温度: 10 °C / 照射時間: 2 sec. / フレーム数: 20 / 単位: 1/A /
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距離分布関数 P(R) | ソフトウェア P(R): GNOM 5.0 / ポイント数: 804 /
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結果 | カーブのタイプ: extrapolated
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