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DNA Origami Scaffolds as Templates for Functional Tetrameric Kir3 K + Channels
Author(s) -
Kurokawa Tatsuki,
Kiyonaka Shigeki,
Nakata Eiji,
Endo Masayuki,
Koyama Shohei,
Mori Emiko,
Tran Nam Ha,
Dinh Huyen,
Suzuki Yuki,
Hidaka Kumi,
Kawata Masaaki,
Sato Chikara,
Sugiyama Hiroshi,
Morii Takashi,
Mori Yasuo
Publication year - 2018
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201709982
Subject(s) - dna origami , template , chemistry , dna , biophysics , scaffold protein , transmembrane protein , nanotechnology , atomic force microscopy , scaffold , zinc finger , biochemistry , biology , materials science , signal transduction , gene , computer science , receptor , database , transcription factor
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.