Versatile in vitro system to study translocation and functional integration of bacterial outer membrane proteins
Versatile in vitro system to study translocation and functional integration of bacterial outer membrane proteins
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DOI：
10.1038/ncomms6396
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发表时间：
2014-11
影响因子：
16.6
通讯作者：
Derrick Norell;A. Heuck;T. Tran‐Thi;H. Götzke;F. Jacob-Dubuisson;T. Clausen;D. Daley;V. Braun;Matthias Müller;Enguo Fan
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文献类型：
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作者：
Derrick Norell;A. Heuck;T. Tran‐Thi;H. Götzke;F. Jacob-Dubuisson;T. Clausen;D. Daley;V. Braun;Matthias Müller;Enguo Fan
革兰氏阴性菌使用V型分泌途径暴露其细胞表面的蛋白质，其中许多具有毒力功能。这些蛋白质跨外膜的转运通过专用的转运蛋白（双伴侣分泌）或共价融合的转运结构域（自身转运蛋白）发生。转运蛋白和转运结构域是β-桶，需要β-桶组装机器（BAM）进行膜整合。然而，它们穿过包膜并插入外膜的分子细节仍然是个谜，部分原因是由于体外系统不可用。在这里，我们描述了一个拟南芥素的体外重建系统，忠实地复制两个分支的V型分泌途径和装配的β桶外膜蛋白。该系统将允许深入分析跨外膜的蛋白质分泌和整合。
Gram-negative bacteria use the type-V secretion pathway to expose proteins at their cell surface, many of which have virulence functions. Translocation of those proteins across the outer membrane occurs either by means of dedicated translocator proteins (two-partner secretion) or covalently fused translocator domains (autotransporters). Translocator proteins and translocator domains are β-barrels requiring the β-barrel assembly machinery (BAM) for membrane integration. However, the molecular details of their passage across the envelope and insertion into the outer membrane remain enigmatic, owing in part to the fact thatin vitrosystems are not available. Here we describe a versatilein vitroreconstitution system that faithfully reproduces both branches of the type-V secretion pathway and the assembly of β-barrel outer membrane proteins. This system will allow an in-depth analysis of protein secretion across and integration into outer membranes.