Encapsulating a single G-quadruplex aptamer in a protein nanocavity.
The alpha-hemolysin (alphaHL) protein pore has many applications in biotechnology. This article describes a single-molecule manipulation system that utilizes the nanocavity enclosed by this pore to noncovalently encapsulate a guest molecule. The guest is the thrombin-binding aptamer (TBA) that folds into the G-quadruplex in the presence of cations. Trapping the G-quadruplex in the nanocavity resulted in characteristic changes to the pore conductance that revealed important molecular processes, including spontaneous unfolding of the quartet structure and translocation of unfolded DNA in the pore. Through detection with Tag-TBA, we localized the G-quadruplex near the entry of the beta-barrel inside the nanocavity, where the molecule vibrates and rotates to different orientations. This guest-nanocavity supramolecular system has potential for helping to understand single-molecule folding and unfolding kinetics.
Department of Biological Engineering and Dalton Cardiovascular Research Center University of Missouri, Columbia, Missouri 65211, USA.
Ji Wook
JW
Shim
Li-Qun
LQ
Gu
D001483:Base Sequence; D054856:G-Quadruplexes; D007700:Kinetics; D049329:Nanostructures / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D014732:Vibration
D009841:Oligonucleotides; D011506:Proteins