1. DNA Origami Scaffolds as Templates for Functional Tetrameric Kir3 K+ Channels
- Author
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Yasuo Mori, Nam Ha Tran, Yuki Suzuki, Hiroshi Sugiyama, Masayuki Endo, Kumi Hidaka, Takashi Morii, Emiko Mori, Masaaki Kawata, Shohei Koyama, Shigeki Kiyonaka, Tatsuki Kurokawa, Huyen Dinh, Eiji Nakata, and Chikara Sato
- Subjects
0301 basic medicine ,Scaffold protein ,Gel electrophoresis ,010405 organic chemistry ,Chemistry ,HEK 293 cells ,General Medicine ,General Chemistry ,010402 general chemistry ,01 natural sciences ,Catalysis ,Transmembrane protein ,0104 chemical sciences ,03 medical and health sciences ,030104 developmental biology ,Template ,Membrane protein ,Biophysics ,DNA origami ,Binding site - 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.
- Published
- 2018