Title
Single-Molecule Fingerprinting of Protein-Drug Interaction Using a Funneled Biological Nanopore
Author
Jeong, Ki-Baek, Minju Ryu, Jin-Sik Kim, Minsoo Kim, Jejoong Yoo, Minji Chung, Sohee Oh, Gyunghee Jo, Seong-Gyu Lee, Ho Min Kim, Mi-Kyung Lee, and Seung-Wook Chi
Year
2023
Journal
Nature Communications
Abstract
In drug discovery, efficient screening of protein-drug interactions (PDIs) is hampered by the limitations of current biophysical approaches. Here, we develop a biological nanopore sensor for single-molecule detection of proteins and PDIs using the pore-forming toxin YaxAB. Using this YaxAB nanopore, we demonstrate label-free, single-molecule detection of interactions between the anticancer Bcl-xL protein and small-molecule drugs as well as the Bak-BH3 peptide. The long funnel-shaped structure and nanofluidic characteristics of the YaxAB nanopore enable the electro-osmotic trapping of diverse folded proteins and high-resolution monitoring of PDIs. Distinctive nanopore event distributions observed in the two-dimensional (ΔI/Io-versus-IN) plot illustrate the ability of the YaxAB nanopore to discriminate individual small-molecule drugs bound to Bcl-xL from non-binders. Taken together, our results present the YaxAB nanopore as a robust platform for label-free, ultrasensitive, single-molecule detection of PDIs, opening up a possibility for low-cost, highly efficient drug discovery against diverse drug targets.
Full Article
Instrument
J-815
Keywords
Biological nanopore sensor