Application of Mistic to improving the expression and membrane integration of histidine kinase receptors from Escherichia coli
Application of Mistic to improving the expression and membrane integration of histidine kinase receptors from Escherichia coli
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DOI：
10.1007/s10969-007-9033-4
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发表时间：
2007-12-01
期刊：
影响因子：
--
通讯作者：
Choe, Senyon
中科院分区：
文献类型：
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作者：
Kefala, Georgia;Kwiatkowski, Witek;Choe, Senyon
整体膜蛋白已成为许多实验室和结构基因组学研究的热点，但其研究在制备、溶解、纯化和结晶等方面存在瓶颈。在我们的实验室，我们已经解决了高水平的蛋白表达的问题，在大肠杆菌的膜使用Mistic，一种新的枯草芽孢杆菌蛋白，作为融合伙伴。在这项研究中，我们研究了Mistic对大肠杆菌组氨酸激酶受体家族成员的蛋白表达和膜整合水平的影响。我们发现Mistic融合总是通过更有效地将货物蛋白靶向到膜上而增加总体产量，甚至可能取代信号序列。Mistic融合方法可能有助于其他整体膜蛋白的高水平表达。
Integral membrane proteins have become the focus of interest of many laboratories and structural genomics consortia, but their study is hampered by bottlenecks in production, solubilization, purification and crystallization. In our laboratory we have addressed the problem of high-level protein expression in the membrane of Escherichia coli by use of Mistic, a novel Bacillus subtilis protein, as a fusion partner. In this study we examine the effect of Mistic on protein expression and membrane integration levels of members of the E. coli histidine kinase receptor family. We find that Mistic fusion invariably increases the overall yield by targeting the cargo proteins more efficiently to the membrane and may even replace the signal sequence. Mistic fusion methods will likely be instrumental for high-level expression of other integral membrane proteins.