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-基本情報
登録情報 | データベース: SASBDB / ID: SASDDC5 |
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試料 | MsbA in stealth nanodisc (SANS, 100% D2O) + 1 mM ADP
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機能・相同性 | 機能・相同性情報 MsbA transporter complex / lipopolysaccharide floppase activity / lipid translocation / ABC-type lipid A-core oligosaccharide transporter / lipopolysaccharide transport / ATPase-coupled lipid transmembrane transporter activity / ABC-type xenobiotic transporter activity / lipid transport / ATP-binding cassette (ABC) transporter complex / transmembrane transport ...MsbA transporter complex / lipopolysaccharide floppase activity / lipid translocation / ABC-type lipid A-core oligosaccharide transporter / lipopolysaccharide transport / ATPase-coupled lipid transmembrane transporter activity / ABC-type xenobiotic transporter activity / lipid transport / ATP-binding cassette (ABC) transporter complex / transmembrane transport / lipid binding / ATP hydrolysis activity / ATP binding / 生体膜 / identical protein binding / 細胞膜 類似検索 - 分子機能 |
生物種 | Escherichia coli (strain K12) (大腸菌) Escherichia coli. (AL95 strain) |
引用 | ジャーナル: Structure / 年: 2018 タイトル: Conformational States of ABC Transporter MsbA in a Lipid Environment Investigated by Small-Angle Scattering Using Stealth Carrier Nanodiscs. 著者: Inokentijs Josts / Julius Nitsche / Selma Maric / Haydyn D Mertens / Martine Moulin / Michael Haertlein / Sylvain Prevost / Dmitri I Svergun / Sebastian Busch / V Trevor Forsyth / Henning Tidow / 要旨: Structural studies of integral membrane proteins (IMPs) are challenging, as many of them are inactive or insoluble in the absence of a lipid environment. Here, we describe an approach making use of ...Structural studies of integral membrane proteins (IMPs) are challenging, as many of them are inactive or insoluble in the absence of a lipid environment. Here, we describe an approach making use of fractionally deuterium labeled "stealth carrier" nanodiscs that are effectively invisible to low-resolution neutron diffraction and enable structural studies of IMPs in a lipidic native-like solution environment. We illustrate the potential of the method in a joint small-angle neutron scattering (SANS) and X-ray scattering (SAXS) study of the ATP-binding cassette (ABC) transporter protein MsbA solubilized in the stealth nanodiscs. The data allow for a direct observation of the signal from the solubilized protein without contribution from the surrounding lipid nanodisc. Not only the overall shape but also differences between conformational states of MsbA can be reliably detected from the scattering data, demonstrating the sensitivity of the approach and its general applicability to structural studies of IMPs. |
登録者 |
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-モデル
-試料
試料 | 名称: MsbA in stealth nanodisc (SANS, 100% D2O) + 1 mM ADP Contrast: -3.313 / Specific vol: 0.745 / 試料濃度: 1.00-6.50 / 濃度測定法: A280 / Entity id: 1016 / 1017 / 1018 |
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バッファ | 名称: 30 mM Tris, 150 mM NaCl, 1 mM ADP / pH: 7.5 / コメント: nanodisc buffer |
要素 #1016 | 名称: MsbA / タイプ: protein / 記述: Lipid A export ATP-binding/permease protein MsbA / 分子量: 66.279 / 分子数: 2 / 由来: Escherichia coli (strain K12) / 参照: UniProt: P60752 配列: MHHHHHHGEN LYFQGSHNDK DLSTWQTFRR LWPTIAPFKA GLIVAGVALI LNAASDTFML SLLKPLLDDG FGKTDRSVLV WMPLVVIGLM ILRGITSYVS SYCISWVSGK VVMTMRRRLF GHMMGMPVSF FDKQSTGTLL SRITYDSEQV ASSSSGALIT VVREGASIIG ...配列: MHHHHHHGEN LYFQGSHNDK DLSTWQTFRR LWPTIAPFKA GLIVAGVALI LNAASDTFML SLLKPLLDDG FGKTDRSVLV WMPLVVIGLM ILRGITSYVS SYCISWVSGK VVMTMRRRLF GHMMGMPVSF FDKQSTGTLL SRITYDSEQV ASSSSGALIT VVREGASIIG LFIMMFYYSW QLSIILIVLA PIVSIAIRVV SKRFRNISKN MQNTMGQVTT SAEQMLKGHK EVLIFGGQEV ETKRFDKVSN RMRLQGMKMV SASSISDPII QLIASLALAF VLYAASFPSV MDSLTAGTIT VVFSSMIALM RPLKSLTNVN AQFQRGMAAC QTLFTILDSE QEKDEGKRVI ERATGDVEFR NVTFTYPGRD VPALRNINLK IPAGKTVALV GRSGSGKSTI ASLITRFYDI DEGEILMDGH DLREYTLASL RNQVALVSQN VHLFNDTVAN NIAYARTEQY SREQIEEAAR MAYAMDFINK MDNGLDTVIG ENGVLLSGGQ RQRIAIARAL LRDSPILILD EATSALDTES ERAIQAALDE LQKNRTSLVI AHRLSTIEKA DEIVVVEDGV IVERGTHNDL LEHRGVYAQL HKMQFGQ |
要素 #1017 | 名称: d75-msp1d1 / タイプ: protein / 記述: Membrane scaffold protein 1D1 (deuterated, 75%) / 分子量: 24.661 / 分子数: 2 配列: GHHHHHHHDY DIPTTENLYF QGSTFSKLRE QLGPVTQEFW DNLEKETEGL RQEMSKDLEE VKAKVQPYLD DFQKKWQEEM ELYRQKVEPL RAELQEGARQ KLHELQEKLS PLGEEMRDRA RAHVDALRTH LAPYSDELRQ RLAARLEALK ENGGARLAEY HAKATEHLST ...配列: GHHHHHHHDY DIPTTENLYF QGSTFSKLRE QLGPVTQEFW DNLEKETEGL RQEMSKDLEE VKAKVQPYLD DFQKKWQEEM ELYRQKVEPL RAELQEGARQ KLHELQEKLS PLGEEMRDRA RAHVDALRTH LAPYSDELRQ RLAARLEALK ENGGARLAEY HAKATEHLST LSEKAKPALE DLRQGLLPVL ESFKVSFLSA LEEYTKKLNT Q |
要素 #1018 | 名称: d(78,92)-POPC / タイプ: other 記述: 1-palmitoyl-2-palmitoleoyl-sn-glycero-3-phosphocholine (deuteration: 78% head, 92% acyl) 分子量: 0.76 / 由来: Escherichia coli. (AL95 strain) 配列: C42H82NO8P |
-実験情報
ビーム | 設備名称: ILL D11 / 地域: Grenoble / 国: France / 線源: neutron source / 波長: 0.46 Å | ||||||||||||||||||||||||||||||||||||||||||
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検出器 | 名称: CERCA / タイプ: 3-He gas detector / Pixsize x: 7.5 mm | ||||||||||||||||||||||||||||||||||||||||||
スキャン | タイトル: (100% D2O) MsbA in stealth nanodisc + 1mM ADP / 測定日: 2017年3月9日 / 保管温度: 10 °C / セル温度: 10 °C / 照射時間: 4200 sec. / フレーム数: 2 / 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: 98 /
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結果 | カーブのタイプ: single_conc / Standard: water コメント: 'MsbA ABC transporter protein encapsulated in a partially deuterated
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