title: Complementary segmental labeling of large RNAs: economic preparation and simplified NMR spectra for measurement of more RDCs.
abstract: NMR structure determination of large RNAs is often restricted by limited RDC information caused by chemical shift degeneracy. We established a general, time- and cost-effective methodology for the preparation of 13C/15N complementary labeled RNAs from a single plasmid. Applying this method to the 25 kDa BC1-DTE RNA, we were able to resolve severe chemical shift degeneracy, thereby almost doubling the number of RDC restraints in comparison to the conventional 13C,15N uniform-labeled RNA.
authors.affiliation: MRC, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom.
authors.forename: Andreas G
authors.initials: AG
authors.lastname: Tzakos
authors.forename: Laura E
authors.initials: LE
authors.lastname: Easton
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Lukavsky
mesh_terms: D002247:Carbon Isotopes; D007097:Imines / Q000737:chemistry; D007553:Isotope Labeling / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009587:Nitrogen Isotopes; D009690:Nucleic Acid Conformation; D004364:Pharmaceutical Preparations / Q000138:chemical synthesis* / Q000191:economics*; D012313:RNA / Q000737:chemistry*; D014158:Transcription, Genetic
chemical_list: D002247:Carbon Isotopes; D007097:Imines; D009587:Nitrogen Isotopes; D004364:Pharmaceutical Preparations; D012313:RNA