Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 57, Issue 10, Pages 2586-2591Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201709982
Keywords
biosensors; DNA origami; DNA structures; ion channels; scaffolds
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Funding
- Ministry of Education, Culture, Sports, Science and Technology (Japan) [24249017, 26111004, 15H01402, 17H01213, 26840032]
- Grants-in-Aid for Scientific Research [17H05440, 16K08494, 15H01402, 16K14033, 15K14499, 16H01562, 15H03837, 17H01213, 26840032] Funding Source: KAKEN
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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 invitro and in living cells.
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