Chemistry Research Laboratory, University of Oxford, Oxford, UK.
Institute for Molecules and Materials, Radboud University, Nijmegen, The Netherlands.
Nat Nanotechnol. 2018 Aug;13(8):739-745. doi: 10.1038/s41565-018-0139-6. Epub 2018 May 28.
The assembly of peptides into membrane-spanning nanopores might be promoted by scaffolds to pre-organize the structures. Such scaffolds could enable the construction of uniform pores of various sizes and pores with controlled permutations around a central axis. Here, we show that DNA nanostructures can serve as scaffolds to arrange peptides derived from the octameric polysaccharide transporter Wza to form uniform nanopores in planar lipid bilayers. Our ring-shaped DNA scaffold is assembled from short synthetic oligonucleotides that are connected to Wza peptides through flexible linkers. When scaffolded, the Wza peptides form conducting nanopores of which only octamers are stable and of uniform conductance. Removal of the DNA scaffold by cleavage of the linkers leads to a rapid loss of the nanopores from the lipid bilayer, which shows that the scaffold is essential for their stability. The DNA scaffold also adds functionality to the nanopores by enabling reversible and permanent binding of complementary tagged oligonucleotides near the nanopore entrance.
多肽组装成跨膜纳米孔可能会被支架促进,以预先组织结构。这种支架可以实现各种尺寸的均匀孔以及围绕中心轴的受控排列的孔的构建。在这里,我们表明 DNA 纳米结构可以作为支架,将源自八聚体多糖转运蛋白 Wza 的肽排列成平面脂质双层中的均匀纳米孔。我们的环形 DNA 支架由短的合成寡核苷酸组装而成,这些寡核苷酸通过柔性接头与 Wza 肽连接。支架化后,Wza 肽形成了导电纳米孔,只有八聚体是稳定的,并且具有均匀的电导率。通过切割接头去除 DNA 支架会导致纳米孔从脂质双层中迅速丢失,这表明支架对于它们的稳定性是必不可少的。DNA 支架还通过在纳米孔入口附近实现互补标记寡核苷酸的可逆和永久结合,为纳米孔添加了功能。