SNARE-fusion mediated insertion of membrane proteins into native and artificial membranes
SNARE-fusion mediated insertion of membrane proteins into native and artificial membranes
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DOI：
10.1038/ncomms5303
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发表时间：
2014-07-01
影响因子：
16.6
通讯作者：
von Ballmoos, Christoph
中科院分区：
文献类型：
--
作者：
Nordlund, Gustav;Brzezinski, Peter;von Ballmoos, Christoph
膜蛋白执行营养吸收、ATP 合成或跨膜信号转导等功能。越来越多的报告表明，细胞过程是由这些蛋白质之间受调节的相互作用支撑的。因此，在分子水平上对这些网络进行功能研究需要对相互作用的成分进行共重建。在这里，我们报告了一种基于 SNARE 蛋白的方法，用于将多种膜蛋白掺入具有明确组成的人工膜囊泡中，并将大的水溶性底物递送到这些囊泡中。该方法用于利用纯化成分在体外重建功能齐全的细菌呼吸链。此外，该方法用于将整个 F1F0 ATP 合酶复合物功能性掺入天然细菌膜中，该成分已从基因上去除。这种新颖的方法提供了一种在分子水平上研究膜结合蛋白之间复杂相互作用网络的工具，有望产生对关键细胞功能的功能见解。
Membrane proteins carry out functions such as nutrient uptake, ATP synthesis or transmembrane signal transduction. An increasing number of reports indicate that cellular processes are underpinned by regulated interactions between these proteins. Consequently, functional studies of these networks at a molecular level require co-reconstitution of the interacting components. Here, we report a SNARE protein-based method for incorporation of multiple membrane proteins into artificial membrane vesicles of well-defined composition, and for delivery of large water-soluble substrates into these vesicles. The approach is used for in vitro reconstruction of a fully functional bacterial respiratory chain from purified components. Furthermore, the method is used for functional incorporation of the entire F1F0 ATP synthase complex into native bacterial membranes from which this component had been genetically removed. The novel methodology offers a tool to investigate complex interaction networks between membrane-bound proteins at a molecular level, which is expected to generate functional insights into key cellular functions.