Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system.
Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system.
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DOI：
10.1038/srep18025
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发表时间：
2015-12-15
影响因子：
4.6
通讯作者：
Akiyoshi K
中科院分区：
文献类型：
--
作者：
Niwa T;Sasaki Y;Uemura E;Nakamura S;Akiyama M;Ando M;Sawada S;Mukai SA;Ueda T;Taguchi H;Akiyoshi K
膜蛋白在细胞过程中起着关键作用，是药物发现的关键靶点。然而，由于膜蛋白具有极高的疏水性，在亲水条件下促进不可逆聚集，因此膜蛋白的可靠合成和折叠是需要解决的重要问题。先前的报道表明，在无细胞翻译过程中加入外源性脂质体可以防止蛋白质聚集。系统地研究哪些膜蛋白可以在脂质体翻译过程中从不可逆聚集中拯救出来，这对于理解脂质体的作用以及在制药科学和纳米器件开发的背景下开发膜蛋白工程的应用是有价值的。因此，我们进行了一项全面的研究，以评估脂质体对大肠杆菌中85种易于聚集的膜蛋白的影响，方法是使用一种重组的、化学定义的无细胞翻译系统。统计分析表明，脂质体的存在提高了所研究膜蛋白的溶解度，最终提高了合成蛋白的收率。生物信息学分析揭示了脂质体效应与膜蛋白的理化性质之间的显著相关性。
Membrane proteins play pivotal roles in cellular processes and are key targets for drug discovery. However, the reliable synthesis and folding of membrane proteins are significant problems that need to be addressed owing to their extremely high hydrophobic properties, which promote irreversible aggregation in hydrophilic conditions. Previous reports have suggested that protein aggregation could be prevented by including exogenous liposomes in cell-free translation processes. Systematic studies that identify which membrane proteins can be rescued from irreversible aggregation during translation by liposomes would be valuable in terms of understanding the effects of liposomes and developing applications for membrane protein engineering in the context of pharmaceutical science and nanodevice development. Therefore, we performed a comprehensive study to evaluate the effects of liposomes on 85 aggregation-prone membrane proteins from Escherichia coli by using a reconstituted, chemically defined cell-free translation system. Statistical analyses revealed that the presence of liposomes increased the solubility of >90% of the studied membrane proteins, and ultimately improved the yields of the synthesized proteins. Bioinformatics analyses revealed significant correlations between the liposome effect and the physicochemical properties of the membrane proteins.