Direct preparation of giant proteo-liposomes by in vitro membrane protein synthesis.
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.
Nomura
Shin-Ichiro M
SM
Institute of Biomaterials & Bioengineering, Tokyo Medical & Dental University, Tokyo, Japan.
Kondoh
Satoshi
S
Asayama
Wakiko
W
Asada
Akikazu
A
Nishikawa
Shigemichi
S
Akiyoshi
Kazunari
K
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
D008081:Liposomes; D011506:Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D015786:Cytochromes b5; D013762:Tetrahydrofolate Dehydrogenase