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首页> 外文期刊>Journal of Molecular Biology >Molecular Organization of Soluble Type III Secretion System Sorting Platform Complexes
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Molecular Organization of Soluble Type III Secretion System Sorting Platform Complexes

机译：可溶性III型分泌系统分选平台复合物的分子组织

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摘要

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(C), 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(C)/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. (C) 2019 Elsevier Ltd. All rights reserved.
机译：许多医学相关的革兰阴性菌使用III型分泌系统（T3SS）易位效应蛋白进入宿主对他们的入侵和细胞内生存。位于T3SS的胞质接口的多蛋白复合物，提出了作为通过选择并通过系统靶向用于分泌基板的分选平台。然而，精确化学计量和分选平台组件的三维组织是未知的。在这里，我们重构鼠伤寒沙门氏菌的可溶性络合物排序平台蛋白，包括ATP酶INVC，调节器OrgB的，蛋白质和SPAO最近鉴定的亚基SPAO（C），这是我们显示成为SPAO的溶解度是必不可少的。我们建立域的域相互作用，确定首次通过本地质谱并深入了解的复合物中每个亚基的化学计量到用小角度X射线散射他们的组织。重要的是，我们发现，在溶液中的组件SPAO / SPAO的（C）/ OrgB的/ INVC采用延长的L形构象类似在原位低温电子断层扫描所见的分拣平台荚，提议这种复杂的是核心构建块可以设想组装成更高的低聚物以形成T3SS排序的平台。分拣平台的可溶性复合物的所确定的分子排列提供了重要的见解其体系结构和组件。 （c）2019 Elsevier Ltd.保留所有权利。

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