Direct observation of the uptake of outer membrane proteins by the periplasmic chaperone Skp.
Direct observation of the uptake of outer membrane proteins by the periplasmic chaperone Skp.
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周质伴侣 Skp 对外膜蛋白摄取的直接观察
DOI：
10.1371/journal.pone.0046068
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Zhao XS
中科院分区：
文献类型：
--
作者：
Lyu ZX;Shao Q;Gao YQ;Zhao XS
膜蛋白在水相亚细胞空间中的运输是一个重要且具有挑战性的过程。其分子机制和相关的结构变化知之甚少。周质分子伴侣，如大肠杆菌中的Skp，在革兰氏阴性菌外膜蛋白（OMP）的运输和保护中起关键作用。Skp与OMP相互作用并保护OMP的分子机制仍然是个谜。本文采用实验和分子动力学模拟相结合的方法，研究了OMPs与Skp结合过程中的结构和动力学信息。停流实验的网站特定的突变和标记Skp和几个外膜蛋白，即OmpC，跨膜结构域的OmpA，和OmpF，使我们能够获得的机制外膜蛋白进入Skp腔，和分子动力学模拟产生详细的分子相互作用负责这一过程。实验和模拟结果均表明，OMP进入Skp是一个高度定向的过程，它是由OMP的N-末端与Skp底部的“tengland”结构域相互作用引发的。Skp的更柔性的张力的打开、OMP和Skp之间的非特异性静电相互作用以及Skp与其底物之间的盐桥的不断形成和断裂一起允许OMP进入Skp并且在没有外部能量供应的情况下逐渐“爬”入Skp腔。
The transportation of membrane proteins through the aqueous subcellular space is an important and challenging process. Its molecular mechanism and the associated structural change are poorly understood. Periplasmic chaperones, such as Skp in Escherichia coli, play key roles in the transportation and protection of outer membrane proteins (OMPs) in Gram-negative bacteria. The molecular mechanism through which Skp interacts with and protects OMPs remains mysterious. Here, a combined experimental and molecular dynamics simulation study was performed to gain the structural and dynamical information in the process of OMPs and Skp binding. Stopped-flow experiments on site specific mutated and labeled Skp and several OMPs, namely OmpC, the transmembrane domain of OmpA, and OmpF, allowed us to obtain the mechanism of OMP entering the Skp cavity, and molecular dynamics simulations yielded detailed molecular interactions responsible for this process. Both experiment and simulation show that the entrance of OMP into Skp is a highly directional process, which is initiated by the interaction between the N-terminus of OMP and the bottom “tentacle” domain of Skp. The opening of the more flexible tentacle of Skp, the non-specific electrostatic interactions between OMP and Skp, and the constant formation and breaking of salt bridges between Skp and its substrate together allow OMP to enter Skp and gradually “climb” into the Skp cavity in the absence of an external energy supply.
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影响因子：
5.6
作者：
Basle, Arnaud;Rummel, Gabriele;Schirmer, Tilman
通讯作者：
Schirmer, Tilman
DOI：
10.1073/pnas.0500127102
发表时间：
2005-02-15
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作者：
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