title: Preparation of RNA samples with narrow line widths for solid state NMR investigations.
abstract: 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 (13)C signals in TAR from >6 ppm to about 1 ppm, making solid state 2D NMR studies of selectively enriched RNAs feasible at ambient temperature.
authors.affiliation: Department of Chemistry, University of Washington, Box 351700, Seattle, WA 98195, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.forename: Michael F
authors.initials: MF
authors.lastname: Bardaro
authors.forename: Gabriele
authors.initials: G
authors.lastname: Varani
authors.forename: Gary P
authors.initials: GP
authors.lastname: Drobny
mesh_terms: D002247:Carbon Isotopes; D015497:HIV-1 / Q000737:chemistry / Q000235:genetics; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011092:Polyethylene Glycols / Q000737:chemistry; D012367:RNA, Viral / Q000737:chemistry / Q000302:isolation & purification*; D015533:Transcriptional Activation
chemical_list: D002247:Carbon Isotopes; D012367:RNA, Viral; D011092:Polyethylene Glycols