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Direct preparation of giant proteo-liposomes by in vitro membrane protein synthesis

✍ Scribed by Shin-ichiro M. Nomura; Satoshi Kondoh; Wakiko Asayama; Akikazu Asada; Shigemichi. Nishikawa; Kazunari Akiyoshi

Book ID
118472522
Publisher
Elsevier Science
Year
2008
Tongue
English
Weight
560 KB
Volume
133
Category
Article
ISSN
0168-1656

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✦ Synopsis

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.

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Liposomes are widely utilized in molecular biology and medicine as drug carriers. Here we report a new liposome-cell interaction through connexins. Connexin 43 (Cx43)-containing liposomes were prepared by using cell-free transcription/translation systems with plasmids encoding Cx43 in the presence o