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-基本情報
登録情報 | データベース: SASBDB / ID: SASDEK7 |
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試料 | SpaO C-terminus
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機能・相同性 | Type III secretion system apparatus protein YscQ/HrcQ/SpaO / Type III secretion system outer membrane, SpaO / Flagellar motor switch protein FliN-like, C-terminal domain / SpoA-like superfamily / Type III flagellar switch regulator (C-ring) FliN C-term / bacterial-type flagellum-dependent swarming motility / protein secretion by the type III secretion system / positive chemotaxis / Surface presentation of antigens protein SpaO 機能・相同性情報 |
生物種 | Salmonella enterica subsp. enterica serovar Typhimurium (サルモネラ菌) |
引用 | ジャーナル: J Mol Biol / 年: 2019 タイトル: Molecular Organization of Soluble Type III Secretion System Sorting Platform Complexes. 著者: Ivonne Bernal / Jonathan Börnicke / Johannes Heidemann / Dmitri Svergun / Julia A Horstmann / Marc Erhardt / Anne Tuukkanen / Charlotte Uetrecht / Michael Kolbe / 要旨: Many medically relevant Gram-negative bacteria use the type III secretion system (T3SS) to translocate effector proteins into the host for their invasion and intracellular survival. A multi-protein ...Many medically relevant Gram-negative bacteria use the type III secretion system (T3SS) to translocate effector proteins into the host for their invasion and intracellular survival. A multi-protein complex located at the cytosolic interface of the T3SS is proposed to act as a sorting platform by selecting and targeting substrates for secretion through the system. However, the precise stoichiometry and 3D organization of the sorting platform components are unknown. Here we reconstitute soluble complexes of the Salmonella Typhimurium sorting platform proteins including the ATPase InvC, the regulator OrgB, the protein SpaO and a recently identified subunit SpaO, which we show to be essential for the solubility of SpaO. We establish domain-domain interactions, determine for the first time the stoichiometry of each subunit within the complexes by native mass spectrometry and gain insight into their organization using small-angle X-ray scattering. Importantly, we find that in solution the assembly of SpaO/SpaO/OrgB/InvC adopts an extended L-shaped conformation resembling the sorting platform pods seen in in situ cryo-electron tomography, proposing that this complex is the core building block that can be conceivably assembled into higher oligomers to form the T3SS sorting platform. The determined molecular arrangements of the soluble complexes of the sorting platform provide important insights into its architecture and assembly. |
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-構造の表示
-ダウンロードとリンク
-モデル
-試料
試料 | 名称: SpaO C-terminus / 試料濃度: 12.5 mg/ml |
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バッファ | 名称: 20 mM HEPES pH 7.5, 150 mM NaCl / pH: 7.5 |
要素 #835 | 名称: SpaO C-terminus / タイプ: protein 記述: Surface presentation of antigens protein SpaO(SPOA1,2) C-terminus 分子量: 19 / 分子数: 1 由来: Salmonella enterica subsp. enterica serovar Typhimurium 参照: UniProt: P40699 配列: MASWSHPQFE KGAVGGGRPK MLRWPLRFVI GSSDTQRSLL GRIGIGDVLL IRTSRAEVYC YAKKLGHFNR VEGGIIVETL DIQHIEEENN TTETAETLPG LNQLPVKLEF VLYRKNVTLA ELEAMGQQQL LSLPTNAELN VEIMANGVLL GNGELVQMND TLGVEIHEWL S |
-実験情報
ビーム | 設備名称: PETRA III EMBL P12 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron / 波長: 0.124 Å / スペクトロメータ・検出器間距離: 3 mm | |||||||||||||||||||||
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検出器 | 名称: Pilatus 2M | |||||||||||||||||||||
スキャン | タイトル: SpaO C-terminus / 測定日: 2017年4月24日 / 保管温度: 20 °C / セル温度: 20 °C / 照射時間: 1 sec. / フレーム数: 3600 / 単位: 1/nm /
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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: 889 /
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結果 | Experimental MW: 20 kDa / カーブのタイプ: sec コメント: SEC-SAXS was performed at 20°C using the following parameters: Column type, XXXX; Flow rate: XXXX mL/min; Total acquisition time: 60 min (3600 x 1 s frames); Injection concentration: ...コメント: SEC-SAXS was performed at 20°C using the following parameters: Column type, XXXX; Flow rate: XXXX mL/min; Total acquisition time: 60 min (3600 x 1 s frames); Injection concentration: XXXX mg/mL; Injection volume: XXXX μL.
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