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American Chemical Society, ACS Nano, 12(5), p. 9345-9353, 2011

DOI: 10.1021/nn203764j

Elsevier, Biophysical Journal, 3(102), p. 429a, 2012

DOI: 10.1016/j.bpj.2011.11.2348

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Nanopore Analysis Of Individual RNA/Antibiotic Complexes

This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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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.