Nanopore Analysis of Individual RNA/Antibiotic Complexes

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• 作者：Meni Wanunu ; Swati Bhattacharya ; Yun Xie ; Yitzhak Tor ; Aleksei Aksimentiev ; Marija Drndic
• 刊名：ACS Nano
• 出版年：2011
• 出版时间：December 27, 2011
• 年：2011
• 卷：5
• 期：12
• 页码：9345-9353
• 全文大小：1016K
• 年卷期：0
• ISSN：1936-086X

文摘

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.