In Vitro Synthesis of the E. coli Sec Translocon from DNA
In Vitro Synthesis of the E. coli Sec Translocon from DNA
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DOI：
10.1002/anie.201403929
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发表时间：
2014-07-14
影响因子：
16.6
通讯作者：
Ueda, Takuya
中科院分区：
文献类型：
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作者：
Matsubayashi, Hideaki;Kuruma, Yutetsu;Ueda, Takuya
构建复杂脂/蛋白膜的困难严重限制了具有重要膜相关功能的功能性人工细胞的发展。通过体外表达其组成蛋白，在脂质囊泡膜中构建了介导膜蛋白向质膜插入的Sec跨位膜通道。Sec转座子的组成部分在脂质体的存在下由各自的基因合成，从而产生一个功能复合物。合成的大肠杆菌Sec易位子介导单跨和多跨膜蛋白的膜易位。功能性肽酶成功易位进入脂质体管腔，进一步证实了易位复合物的正确插入。我们的研究结果证明了构建人工细胞的可行性，通过将遗传信息直接解码为膜功能，可以实现人工细胞膜的功能化。
Difficulties in constructing complex lipid/protein membranes have severely limited the development of functional artificial cells endowed with vital membrane-related functions. The Sec translocon membrane channel, which mediates the insertion of membrane proteins into the plasma membrane, was constructed in the membrane of lipid vesicles through in vitro expression of its component proteins. The components of the Sec translocon were synthesized from their respective genes in the presence of liposomes, thereby bringing about a functional complex. The synthesized E. coli Sec translocon mediated the membrane translocation of single- and multi-span membrane proteins. The successful translocation of a functional peptidase into the liposome lumen further confirmed the proper insertion of the translocon complex. Our results demonstrate the feasible construction of artificial cells, the membranes of which can be functionalized by directly decoding genetic information into membrane functions.