DNA Origami Scaffolds as Templates for Functional Tetrameric Kir3 K+ Channels

Tatsuki Kurokawa, Shigeki Kiyonaka, Eiji Nakata, Masayuki Endo, Shohei Koyama, Emiko Mori, Nam Ha Tran, Huyen Dinh, Yuki Suzuki, Kumi Hidaka, Masaaki Kawata, Chikara Sato, Hiroshi Sugiyama, Takashi Morii, Yasuo Mori

Research output: Contribution to journalArticle

13 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)2586-2591
Number of pages6
JournalAngewandte Chemie - International Edition
Volume57
Issue number10
DOIs
Publication statusPublished - 2018 Mar 1
Externally publishedYes

Keywords

  • DNA origami
  • DNA structures
  • biosensors
  • ion channels
  • scaffolds

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)

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    Kurokawa, T., Kiyonaka, S., Nakata, E., Endo, M., Koyama, S., Mori, E., Tran, N. H., Dinh, H., Suzuki, Y., Hidaka, K., Kawata, M., Sato, C., Sugiyama, H., Morii, T., & Mori, Y. (2018). DNA Origami Scaffolds as Templates for Functional Tetrameric Kir3 K+ Channels. Angewandte Chemie - International Edition, 57(10), 2586-2591. https://doi.org/10.1002/anie.201709982