| Affiliation: | 1. Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyotodaigakukatsura, Nishikyo-ku, Kyoto, Japan;2. Core Research for Evolutional Science and Technology (Japan), Science and Technology Agency, Saitama, Japan;3. Department of Technology and Ecology, Hall of Global Environmental Studies, Kyoto University, Kyotodaigakukatsura, Nishikyo-ku, Kyoto, Japan;4. Institute of Advanced Energy, Kyoto University, Gokasho, Uji, Kyoto, Japan;5. Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida-Ushinomiya-cho, Sakyo-ku, Kyoto, Japan;6. Department of Chemistry, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake-cho, Sakyo-ku, Kyoto, Japan;7. Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Ibaraki, Japan |
| Abstract: | In native systems, scaffolding proteins play important roles in assembling proteins into complexes to transduce signals. This concept is yet to be applied to the assembly of functional transmembrane protein complexes in artificial systems. To address this issue, DNA origami has the potential to serve as scaffolds that arrange proteins at specific positions in complexes. Herein, we report that Kir3 K+ channel proteins are assembled through zinc‐finger protein (ZFP)‐adaptors at specific locations on DNA origami scaffolds. Specific binding of the ZFP‐fused Kir3 channels and ZFP‐based adaptors on DNA origami were confirmed by atomic force microscopy and gel electrophoresis. Furthermore, the DNA origami with ZFP binding sites nearly tripled the K+ channel current activity elicited by heterotetrameric Kir3 channels in HEK293T cells. Thus, our method provides a useful template to control the oligomerization states of membrane protein complexes in vitro and in living cells. |
