Digital nanoreactors to control absolute stoichiometry and spatiotemporal behavior of DNA receptors within lipid bilayers

Vishal Maingi (), Zhao Zhang, Chris Thachuk (), Namita Sarraf, Edwin R. Chapman () and Paul W. K. Rothemund ()
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Vishal Maingi: Department of Bioengineering, California Institute of Technology
Zhao Zhang: University of Wisconsin-Madison
Chris Thachuk: University of Washington
Namita Sarraf: Department of Bioengineering, California Institute of Technology
Edwin R. Chapman: University of Wisconsin-Madison
Paul W. K. Rothemund: Department of Bioengineering, California Institute of Technology

Nature Communications, 2023, vol. 14, issue 1, 1-12

Abstract: Abstract Interactions between membrane proteins are essential for cell survival but are often poorly understood. Even the biologically functional ratio of components within a multi-subunit membrane complex—the native stoichiometry—is difficult to establish. Here we demonstrate digital nanoreactors that can control interactions between lipid-bound molecular receptors along three key dimensions: stoichiometric, spatial, and temporal. Each nanoreactor is based on a DNA origami ring, which both templates the synthesis of a liposome and provides tethering sites for DNA-based receptors (modelling membrane proteins). Receptors are released into the liposomal membrane using strand displacement and a DNA logic gate measures receptor heterodimer formation. High-efficiency tethering of receptors enables the kinetics of receptors in 1:1 and 2:2 absolute stoichiometries to be observed by bulk fluorescence, which in principle is generalizable to any ratio. Similar single-molecule-in-bulk experiments using DNA-linked membrane proteins could determine native stoichiometry and the kinetics of membrane protein interactions for applications ranging from signalling research to drug discovery.

Date: 2023
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DOI: 10.1038/s41467-023-36996-x

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