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基本情報
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試料 | The InvC-ATP synthase, surface presentation of antigens and oxygen-regulated invasion protein (ISO) complex
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| 機能・相同性 | 機能・相同性情報protein-secreting ATPase / type III protein secretion system complex / protein secretion by the type III secretion system / bacterial-type flagellum-dependent swarming motility / positive chemotaxis / proton-transporting ATPase activity, rotational mechanism / proton-transporting ATP synthase activity, rotational mechanism / protein transport / ATP hydrolysis activity / ATP binding / cytoplasm 類似検索 - 分子機能 |
| 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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試料
試料 | 名称: The InvC-ATP synthase, surface presentation of antigens and oxygen-regulated invasion protein (ISO) complex 試料濃度: 1 mg/ml / Entity id: 824 / 825 / 1359 / 1360 |
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| バッファ | 名称: 10 mM Tris-HCl, 50 mM NaCl / pH: 8 |
| 要素 #824 | 名称: SpaOc / タイプ: protein 記述: Surface presentation of antigens protein SpaO SpaO(SPOA2) 分子量: 12.374 / 分子数: 2 由来: Salmonella enterica subsp. enterica serovar Typhimurium 参照: UniProt: P40699 配列: METLDIQHIE EENNTTETAE TLPGLNQLPV KLEFVLYRKN VTLAELEAMG QQQLLSLPTN AELNVEIMAN GVLLGNGELV QMNDTLGVEI HEWLSESGNG ESAWSHPQFE K |
| 要素 #825 | 名称: SpaO / タイプ: protein 記述: Surface presentation of antigens protein SpaO(SPOA1,2) 分子量: 33.793 / 分子数: 1 由来: Salmonella enterica subsp. enterica serovar Typhimurium 参照: UniProt: P40699 配列: MSLRVRQIDR REWLLAQTAT ECQRHGREAT LEYPTRQGMW VRLSDAEKRW SAWIKPGDWL EHVSPALAGA AVSAGAEHLV VPWLAATERP FELPVPHLSC RRLCVENPVP GSALPEGKLL HIMSDRGGLW FEHLPELPAV GGGRPKMLRW PLRFVIGSSD TQRSLLGRIG ...配列: MSLRVRQIDR REWLLAQTAT ECQRHGREAT LEYPTRQGMW VRLSDAEKRW SAWIKPGDWL EHVSPALAGA AVSAGAEHLV VPWLAATERP FELPVPHLSC RRLCVENPVP GSALPEGKLL HIMSDRGGLW FEHLPELPAV GGGRPKMLRW PLRFVIGSSD TQRSLLGRIG IGDVLLIRTS RAEVYCYAKK LGHFNRVEGG IIVETLDIQH IEEENNTTET AETLPGLNQL PVKLEFVLYR KNVTLAELEA MGQQQLLSLP TNAELNVEIM ANGVLLGNGE LVQMNDTLGV EIHEWLSESG NGE |
| 要素 #1359 | 名称: OrgB / タイプ: protein / 記述: Oxygen-regulated invasion protein OrgB / 分子量: 26.46 / 分子数: 2 由来: Salmonella enterica subsp. enterica serovar Typhimurium 参照: UniProt: P0CL45 配列: MLKNIPIPSP LSPVEGILIK RKTLERYFSI ERLEQQAHQR AKRILREAEE EAKTLRMYAY QEGYEQGMID ALQQVAAYLT DNQTMAWKWM EKIQIYAREL FSAAVDHPET LLTVLDEWLR DFDKPEGQLF LTLPVNAKKD HQKLMVLLME NWPGTFNLKY HQEQRFIMSC ...配列: MLKNIPIPSP LSPVEGILIK RKTLERYFSI ERLEQQAHQR AKRILREAEE EAKTLRMYAY QEGYEQGMID ALQQVAAYLT DNQTMAWKWM EKIQIYAREL FSAAVDHPET LLTVLDEWLR DFDKPEGQLF LTLPVNAKKD HQKLMVLLME NWPGTFNLKY HQEQRFIMSC GDQIAEFSPE QFVETAVGVI KHHLDELPQD CRTISDNAIN ALIDEWKTKT QAEVIR |
| 要素 #1360 | 名称: InvC / タイプ: protein / 記述: ATP synthase InvC / 分子量: 47.61 / 分子数: 1 由来: Salmonella enterica subsp. enterica serovar Typhimurium 参照: UniProt: E8XL22 配列: MKTPRLLQYL AYPQKITGPI IEAELRDVAI GELCEIRRGW HQKQVVARAQ VVGLQRERTV LSLIGNAQGL SRDVVLYPTG RALSAWVGYS VLGAVLDPTG KIVERFTPEV APISEERVID VAPPSYASRV GVREPLITGV RAIDGLLTCG VGQRMGIFAS AGCGKTMLMH ...配列: MKTPRLLQYL AYPQKITGPI IEAELRDVAI GELCEIRRGW HQKQVVARAQ VVGLQRERTV LSLIGNAQGL SRDVVLYPTG RALSAWVGYS VLGAVLDPTG KIVERFTPEV APISEERVID VAPPSYASRV GVREPLITGV RAIDGLLTCG VGQRMGIFAS AGCGKTMLMH MLIEQTEADV FVIGLIGERG REVTEFVDML RASHKKEKCV LVFATSDFPS VDRCNAAQLA TTVAEYFRDQ GKRVVLFIDS MTRYARALRD VALASGERPA RRGYPASVFD NLPRLLERPG ATSEGSITAF YTVLLESEEE ADPMADEIRS ILDGHLYLSR KLAGQGHYPA IDVLKSVSRV FGQVTTPTHA EQASAVRKLM TRLEELQLFI DLGEYRPGEN IDNDRAMQMR DSLKAWLCQP VAQYSSFDDT LSGMNAFADQ N |
-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 | 測定日: 2017年4月12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) |
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| 結果 | 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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Salmonella enterica subsp. enterica serovar Typhimurium (サルモネラ菌)
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SASDEJ7







