Reconstitution of a nanomachine driving the assembly of proteins into bacterial outer membranes.
Reconstitution of a nanomachine driving the assembly of proteins into bacterial outer membranes.
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DOI：
10.1038/ncomms6078
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发表时间：
2014-10-24
影响因子：
16.6
通讯作者：
Lithgow T
中科院分区：
文献类型：
--
作者：
Shen HH;Leyton DL;Shiota T;Belousoff MJ;Noinaj N;Lu J;Holt SA;Tan K;Selkrig J;Webb CT;Buchanan SK;Martin LL;Lithgow T
在生物膜中，各种蛋白质分泌设备的功能就像纳米机器一样，测量其组成部分的内部运动是一项重大的技术挑战。易位组装模块()是细菌病原体毒力所需的纳米机器。我们用耗散石英晶体微天平(QCM-D)和磁对比度中子反射仪(MCNR)对含有的膜进行了表征。MCNR研究提供了低至1？的结构分辨率，使准确测量从膜层突出的蛋白质结构域成为可能。在这里，我们证明了的TAMA组件内的动态运动是在底物蛋白Ag43存在的情况下启动的，并且这些运动概括了膜蛋白组装的初始阶段。该重组系统为研究生物膜中的分子运动提供了一种强有力的新手段，该技术广泛适用于各种细胞纳米机器的动力学研究。
In biological membranes, various protein secretion devices function as nanomachines, and measuring the internal movements of their component parts is a major technological challenge. The translocation assembly module (the TAM) is a nanomachine required for virulence of bacterial pathogens. We have reconstituted a membrane containing the TAM onto a gold surface for characterization by Quartz Crystal Microbalance with Dissipation (QCM-D) and Magnetic Contrast Neutron Reflectrometry (MCNR). The MCNR studies provided structural resolution down to 1Å, enabling accurate measurement of protein domains projecting from the membrane layer. Here, we show that dynamic movements within the TamA component of the TAM are initiated in the presence of a substrate protein, Ag43, and that these movements recapitulate an initial stage in membrane protein assembly. The reconstituted system provides a powerful new means to study molecular movements in biological membranes, and the technology is widely applicable to studying the dynamics of diverse cellular nanomachines.