Single-molecule bonds characterized by solid-state nanopore force spectroscopy.
Weak molecular interactions drive processes at the core of living systems, such as enzyme-substrate interactions, receptor-ligand binding, and nucleic acid replication. Single-molecule force spectroscopy is a remarkable tool for revealing molecular scale energy landscapes of noncovalent bonds, by exerting a mechanical force directly on an individual molecular complex and tracking its survival as a function of time and applied force. In principle, force spectroscopy methods can also be used for highly specific molecular recognition assays, by directly characterizing the strength of bonds between probe and target molecules. However, complexity and low throughput of conventional force spectroscopy techniques render such biosensing applications impractical. Here we demonstrate a straightforward single-molecule approach, suitable for both biophysical studies and molecular recognition assays, in which a approximately 3 nm silicon nitride nanopore is used to determine the bond lifetime spectrum of the biotin-neutravidin complex. Thousands of individual molecular complexes are captured and dissociated in the solid-state nanopore under constant applied forces, ranging from 400 to 900 mV, allowing us to extract the location of the energy barrier that governs the interaction, mapped at Deltax approximately 0.5 nm. These results highlight the capacity of a solid-state nanopore to detect and characterize intermolecular interactions and demonstrate how this could be applied to rapid, highly specific molecular detection assays.
Tabard-Cossa
Vincent
V
Department of Physics and Astronomy, University of British Columbia, B.C., Canada.
Wiggin
Matthew
M
Trivedi
Dhruti
D
Jetha
Nahid N
NN
Dwyer
Jason R
JR
Marziali
Andre
A
D001360:Avidin / Q000378:metabolism; D001483:Base Sequence; D001710:Biotin / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism; D008024:Ligands; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000379:methods*; D016062:Porosity; D017655:Silicon Compounds / Q000737:chemistry; D013057:Spectrum Analysis / Q000379:methods*; D013816:Thermodynamics
D008024:Ligands; D017655:Silicon Compounds; C093483:neutravidin; D001360:Avidin; D001710:Biotin; D004247:DNA; C032734:silicon nitride