title: Direct preparation of giant proteo-liposomes by in vitro membrane protein synthesis.
abstract: We investigated the direct constitution of membrane proteins into giant liposomes in cell-free (in vitro) protein synthesis. Giant liposomes were present in a translation reaction cocktail of a wheat germ cell-free protein translation system. Apo cytochrome b(5) (b5) and its fusion proteins were synthesized and directly localized in the liposomes. After the translation reaction, the proteo-liposomes were isolated by simplified discontinuous density-gradient centrifugation. Apo cytochrome b(5) conjugated dihydrofolate reductase (DHFR) was synthesized in the same procedure and the protein was directly displayed on the liposome surface. b5 acts as a "hydrophobic tag" for recruitment to the liposome surface.
authors: Institute of Biomaterials & Bioengineering, Tokyo Medical & Dental University, Tokyo, Japan.
authors: Shin-Ichiro M
authors: SM
authors: Nomura
authors: Satoshi
authors: S
authors: Kondoh
authors: Wakiko
authors: W
authors: Asayama
authors: Akikazu
authors: A
authors: Asada
authors: Shigemichi
authors: S
authors: Nishikawa
authors: Kazunari
authors: K
authors: Akiyoshi
mesh_terms: D015786:Cytochromes b5 / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D014176:Protein Biosynthesis; D011506:Proteins / Q000378:metabolism*; D013762:Tetrahydrofolate Dehydrogenase / Q000378:metabolism; D013997:Time Factors
chemical_list: D008081:Liposomes; D011506:Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D015786:Cytochromes b5; D013762:Tetrahydrofolate Dehydrogenase