Abstract
DNA nanotechnology allows for the creation of three-dimensional structures at nanometer scale. Here, we use DNA to build the largest synthetic pore in a lipid membrane to date, approaching the dimensions of the nuclear pore complex and increasing the pore-area and the conductance 10-fold compared to previous man-made channels. In our design, 19 cholesterol tags anchor a megadalton funnel-shaped DNA origami porin in a lipid bilayer membrane. Confocal imaging and ionic current recordings reveal spontaneous insertion of the DNA porin into the lipid membrane, creating a transmembrane pore of tens of nanosiemens conductance. All-atom molecular dynamics simulations characterize the conductance mechanism at the atomic level and independently confirm the DNA porins’ large ionic conductance.
Highlights
D ue to their diverse architectures, protein channels in natural lipid membranes are capable of fulfilling a variety of functions in living cells, from the recognition of substrates to the selective transport of ions or large biomolecules between cellular compartments.[1]
We expand the design space of synthetic lipid membrane pores beyond these limits by creating a significantly larger funnel-shaped porin from DNA origami.[13]
The nominal cross section of the DNA porin is 6 nm, Figure 1B, which is wider than the cross section of large natural porins[14] and comparable to the electrical diameter of the nuclear pore complex.[15]
Summary
D ue to their diverse architectures, protein channels in natural lipid membranes are capable of fulfilling a variety of functions in living cells, from the recognition of substrates to the selective transport of ions or large biomolecules between cellular compartments.[1] Synthetic channels have been proposed as components of drug-delivery systems, as antimicrobial agents, biosensors, filters, photosystems, catalysts,[2] or as tools for synthetic biology;[3] all undoubtedly requiring tailored architectures with a high level of customizability.
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