Atomic model of the type III secretion system needle.
Atomic model of the type III secretion system needle.
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DOI：
10.1038/nature11079
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发表时间：
2012-05-20
期刊：
影响因子：
64.8
通讯作者：
Lange, Adam
中科院分区：
文献类型：
--
作者：
Loquet, Antoine;Sgourakis, Nikolaos G.;Gupta, Rashmi;Giller, Karin;Riedel, Dietmar;Goosmann, Christian;Griesinger, Christian;Kolbe, Michael;Baker, David;Becker, Stefan;Lange, Adam
使用III型分泌系统（T3 SS）操纵宿主细胞的病原菌引起许多不同的感染，包括志贺氏痢疾、伤寒、肠出血性结肠炎和腺鼠疫。T3 SS的一个重要部分是中空的针状蛋白丝，效应蛋白通过该蛋白丝被注射到真核宿主细胞中。目前，针的三维结构是未知的，因为它不服从X射线晶体学和溶液NMR，由于其固有的非结晶性和不溶性。冷冻电子显微镜结合晶体或溶液NMR亚基结构最近提供了一个强大的混合方法研究超分子组装，导致低分辨率和中等分辨率的模型。然而，这种方法不能提供原子的细节，特别是关键的亚基-亚基界面，因为在这些研究中获得的有限的冷冻电子显微镜分辨率。在这里，我们报告了一种替代方法相结合的重组野生型针生产，固态NMR，电子显微镜和Rosetta建模揭示超分子界面，并最终完整的原子结构的鼠伤寒沙门氏菌T3 SS针。我们发现，80个残基的亚基形成了一个右手螺旋组装，每两圈大约有11个亚基，类似于S的鞭毛丝。鼠伤寒。与已建立的针模型（其中蛋白质亚基的氨基末端被假定为α-螺旋并位于针内）相反，我们的模型揭示了位于针表面的延伸氨基末端结构域，而高度保守的羧基末端指向管腔。
Pathogenic bacteria using a type III secretion system (T3SS) to manipulate host cells cause many different infections including Shigella dysentery, typhoid fever, enterohaemorrhagic colitis and bubonic plague. An essential part of the T3SS is a hollow needle-like protein filament through which effector proteins are injected into eukaryotic host cells. Currently, the three-dimensional structure of the needle is unknown because it is not amenable to X-ray crystallography and solution NMR, as a result of its inherent non-crystallinity and insolubility. Cryo-electron microscopy combined with crystal or solution NMR subunit structures has recently provided a powerful hybrid approach for studying supramolecular assemblies, resulting in low-resolution and medium-resolution models. However, such approaches cannot deliver atomic details, especially of the crucial subunit–subunit interfaces, because of the limited cryo-electron microscopic resolution obtained in these studies. Here we report an alternative approach combining recombinant wild-type needle production, solid-state NMR, electron microscopy and Rosetta modelling to reveal the supramolecular interfaces and ultimately the complete atomic structure of the Salmonella typhimurium T3SS needle. We show that the 80-residue subunits form a right-handed helical assembly with roughly 11 subunits per two turns, similar to that of the flagellar filament of S. typhimurium. In contrast to established models of the needle in which the amino terminus of the protein subunit was assumed to be α-helical and positioned inside the needle, our model reveals an extended amino-terminal domain that is positioned on the surface of the needle, while the highly conserved carboxy terminus points towards the lumen.
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影响因子：
64.8
作者：
Fujii, Takashi;Iwane, Atsuko H.;Namba, Keiichi
通讯作者：
Namba, Keiichi
影响因子：
56.9
作者：
Schraidt, Oliver;Marlovits, Thomas C.
通讯作者：
Marlovits, Thomas C.
DOI：
10.1073/pnas.200209497
发表时间：
2000-09-26
影响因子：
11.1
作者：
Kimbrough, TG;Miller, SI
通讯作者：
Miller, SI
影响因子：
15
作者：
Lewandowski, Jozef R.;De Paepe, Gael;Griffin, Robert G.
通讯作者：
Griffin, Robert G.
影响因子：
15
作者：
Loquet, Antoine;Lv, Guohua;Lange, Adam
通讯作者：
Lange, Adam