Nanopore analysis of individual RNA/antibiotic complexes

ACS Nano. 2011 Dec 27;5(12):9345-53. doi: 10.1021/nn203764j. Epub 2011 Nov 16.

Abstract

Nanopores in thin solid-state membranes are used to rapidly analyze individual RNA/drug complexes. The interactions of a truncated A-site RNA model of the prokaryotic ribosome with aminoglycoside antibiotics are characterized by passing individual molecules through a 3-3.5 nm diameter pore fabricated in a 8-10 nm thick silicon nitride membrane. Complexes of the A-site RNA with aminoglycosides can be distinguished from unbound A-site based on the ion current signatures produced as they pass through the nanopores. Counting the fraction of free and drug-bound molecules affords label-free drug-RNA binding isotherms consistent with literature reports and with data generated using independent fluorescence-based assays. Our measurements are supported by molecular dynamics simulations, which illustrate the relationship between the ionic current and complexation of the A-site RNA with paramomycin, a prototypical aminoglycoside antibiotic.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Anti-Bacterial Agents / chemistry
  • Binding Sites
  • Diffusion
  • Electroporation / methods*
  • Materials Testing
  • Membranes, Artificial*
  • Nanostructures / chemistry*
  • Nanostructures / ultrastructure*
  • Paromomycin / chemistry*
  • Porosity
  • RNA / chemistry*
  • Semiconductors

Substances

  • Anti-Bacterial Agents
  • Membranes, Artificial
  • Paromomycin
  • RNA