title: In vitro synthesis of the E. coli Sec translocon from DNA.
abstract: 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.
authors.affiliation: Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa-shi, 277-8562 Chiba (Japan).
authors.forename: Hideaki
authors.initials: H
authors.lastname: Matsubayashi
authors.forename: Yutetsu
authors.initials: Y
authors.lastname: Kuruma
authors.forename: Takuya
authors.initials: T
authors.lastname: Ueda
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D004269:DNA, Bacterial / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry*; D066298:In Vitro Techniques
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial
keywords: Sec translocon; artificial cells; cell-free systems; membrane proteins; synthetic biology