Preparation of RNA samples with narrow line widths for solid state NMR investigations

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Preparation of RNA samples with narrow line widths for solid state NMR investigations

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08-10-2012, 08:40 PM

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Preparation of RNA samples with narrow line widths for solid state NMR investigations

Preparation of RNA samples with narrow line widths for solid state NMR investigations

Publication year: 2012
Source:Journal of Magnetic Resonance

Wei Huang, Michael F. Bardaro, Gabriele Varani, Gary P. Drobny

Solid state NMR can provide detailed structural and dynamic information on biological systems that cannot be studied under solution conditions, and can investigate motions which occur with rates that cannot be fully studied by solution NMR. This approach has successfully been used to study proteins, but the application of multidimensional solid state NMR to RNA has been limited because reported line widths have been too broad to execute most multidimensional experiments successfully. A reliable method to generate spectra with narrow line widths is necessary to apply the full range of solid state NMR spectroscopic approaches to RNA. Using the HIV-1 transactivation response (TAR) RNA as a model, we present an approach based on precipitation with polyethylene glycol that improves the line width of 13C signals in TAR from >6 ppm to about 1 ppm, making solid state 2D NMR studies of selectively enriched RNAs feasible at ambient temperature.
Graphical abstract

Graphical abstract Highlights

? Narrow line widths are required to apply the full range of solid state NMR to RNA. ? Precipitation with PEG significantly improves the line width of TAR RNA. ? 2D SSNMR studies of selectively enriched RNAs are feasible at ambient temperature.

Source: Journal of Magnetic Resonance

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