Preparation of connexin43-integrated giant Liposomes by a baculovirus expression-liposome fusion method.
Connexin-43 (Cx43) containing giant liposomes (GL) were prepared by a baculovirus expression-liposome fusion method. Recombinant budded viruses expressing Cx43 were prepared and then fused with GLs containing DOPG/DOPC at pH 4.5. Connexon formation on the GL membrane was observed by transmission electron microscope. Hydrophilic fluorescent dye transfers were observed through a Cx43-mediated pathway not only between Sf9 (Spodoptera frugiperda) cells with Cx43 but also from giant Cx43 liposomes to Cx43-expressing U2OS cells (human osteosarcoma cell). The functional connexin-containing liposome is expected to be useful for cellular cytosolic delivery systems. The original orientation and function of Cx43 was maintained after integration into the liposomes. The liposome fusion method will create new opportunities as a tool for analysis of channel membrane proteins.
Kamiya
Koki
K
Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-0062, Japan.
Tsumoto
Kanta
K
Arakawa
Satoko
S
Shimizu
Shigeomi
S
Morita
Ikuo
I
Yoshimura
Tetsuro
T
Akiyoshi
Kazunari
K
D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000254:growth & development*; D001709:Biotechnology / Q000379:methods*; D002460:Cell Line; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D008081:Liposomes / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D018411:Spodoptera; D013194:Staining and Labeling / Q000379:methods; D014771:Virion / Q000378:metabolism*
D018031:Connexin 43; D005456:Fluorescent Dyes; D008081:Liposomes; D011994:Recombinant Proteins