Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa-shi, 277-8562 Chiba (Japan).
Angew Chem Int Ed Engl. 2014 Jul 14;53(29):7535-8. doi: 10.1002/anie.201403929. Epub 2014 Jun 4.
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.
构建复杂的脂质/蛋白质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。Sec 易位子膜通道介导膜蛋白插入质膜，通过在体外表达其组成蛋白在脂质体的膜中构建。Sec 易位子的组成部分在脂质体存在的情况下从各自的基因中合成，从而形成功能性复合物。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。功能肽酶成功转位到脂质体腔中进一步证实了易位子复合物的正确插入。我们的结果表明，可以通过直接将遗传信息解码为膜功能来构建人工细胞，这些人工细胞的膜可以通过功能化。
