TY - JOUR
T1 - Direct preparation of giant proteo-liposomes by in vitro membrane protein synthesis
AU - Nomura, Shin ichiro M.
AU - Kondoh, Satoshi
AU - Asayama, Wakiko
AU - Asada, Akikazu
AU - Nishikawa, Shigemichi
AU - Akiyoshi, Kazunari
N1 - Funding Information:
This research was supported by a grant from the Center of Excellence Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone at Tokyo Medical and Dental University. This work was also supported by Grants-in-Aid for Scientific Research from the Japanese Government (Nos. 18GS0421, 18048013, 18048014). Pacific Edit reviewed the manuscript prior to submission.
PY - 2008/1/20
Y1 - 2008/1/20
N2 - 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 b5 (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 b5 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.
AB - 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 b5 (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 b5 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.
KW - Cytochrome b5
KW - Giant liposome
KW - In vitro protein synthesis
KW - Membrane protein
KW - Wheat germ lysate
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U2 - 10.1016/j.jbiotec.2007.08.023
DO - 10.1016/j.jbiotec.2007.08.023
M3 - Article
C2 - 17900734
AN - SCOPUS:37249051888
SN - 0168-1656
VL - 133
SP - 190
EP - 195
JO - Journal of Biotechnology
JF - Journal of Biotechnology
IS - 2
ER -