Complementary segmental labeling of large RNAs: economic preparation and simplified NMR spectra for measurement of more RDCs.
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.
Tzakos
Andreas G
AG
MRC, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom.
Easton
Laura E
LE
Lukavsky
Peter J
PJ
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
D002247:Carbon Isotopes; D007097:Imines; D009587:Nitrogen Isotopes; D004364:Pharmaceutical Preparations; D012313:RNA