Summary
An approach for the simultaneous acquisition of HCN and HCP as well as HCN-CCH-TOCSY and HCP-CCH-TOCSY triple resonance data sets for 13C-/15N-labelled RNAs is presented. The new HCN-CCH-TOCSY scheme unambiguously links all sugar resonances to the base nitrogen. In addition, simultaneous acquisition of HCN-CCH-TOCSY and HCP-CCH-TOCSY data sets provides sequential and base-type information in a single experiment, thereby saving data acquisition time as well as providing complementary data sets that are useful in clarifying ambiguous assignments. Virtually complete sequence-specific phosphate-ribose 1H, 31P, and base 15N1,9 assignments as well as partial 13C assignments could be obtained in a single experiment for a 0.5-mM sample of a 19-mer ribonucleotide.
Article PDF
Avoid common mistakes on your manuscript.
References
Aboul-elaF. and VaraniG. (1995) Curr. Opin. Biotechnol., 6, 89–95.
BateyR.T., InadaM., KujawinskiE., PuglisiJ.D. and WilliamsonJ.R. (1992) Nucleic Acids Res., 20, 4515–4523.
DieckmannT. and FeigonJ. (1994) Curr. Opin. Struct. Biol., 4, 745–749.
FarmerIIB.T. (1991) J. Magn. Reson., 93, 635–641.
FarmerIIB.T., MuellerL., NikonowiczE.P. and PardiA. (1993) J. Am. Chem. Soc., 115, 11040–11041.
FarmerIIB.T., MuellerL., NikonowiczE.P. and PardiA. (1994) J. Biomol. NMR, 4, 129–133.
HeusH.A., WijmengaS.S., Van deVenF.J.M. and HilbersC.W. (1994) J. Am. Chem. Soc., 116, 4983–4984.
MarinoJ.P., SchwalbeH., AnklinC., BermelW., CrothersD.M. and GriesingerC. (1994) J. Am. Chem. Soc., 116, 6472–6473.
MarinoJ.P., SchwalbeH., AnklinC., BermelW., CrothersD.M. and GriesingerC. (1995) J. Biomol. NMR, 5, 87–92.
NikonowiczE.P., SirrA., LegaultP., JuckerF.M., BaerL.M. and PardiA. (1992) Nucleic Acids Res., 20, 4507–4513.
PardiA. and NikonowiczE.P. (1992) J. Am. Chem. Soc., 114, 9202–9203.
SklenářV., PetersonR.D., RejanteM.R. and FeigonJ. (1993a) J. Biomol. NMR, 3, 721–727.
SklenářV., PetersonR.D., RejanteM.R., WangE. and FeigonJ. (1993b) J. Am. Chem. Soc., 115, 12181–12182.
SklenářV., PetersonR.D., RejanteM.R. and FeigonJ. (1994) J. Biomol. NMR, 4, 117–122.
SørensenO.W. (1990) J. Magn. Reson., 89, 210–216.
TateS.I., OnoA. and KainoshoM. (1994) J. Am. Chem. Soc., 116, 5977–5978.
TateS.I., OnoA. and KainoshoM. (1995) J. Magn. Reson. Ser. B, 106, 89–91.
VaraniG. and TinocoJr.I. (1991) Q. Rev. Biophys., 24, 479–532.
WijmengaS.S., HeusH.A., LeeuwH.A.E., HoppeH., Van derGraafM. and HilbersC.W. (1995) J. Biomol. NMR, 5, 82–86.
WoltersJ. (1992) Nucleic Acids Res., 20, 1843–1850.
WyattJ.R., ChastainM. and PuglisiJ.D. (1991) Biotechniques, 11, 764–769.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ramachandran, R., Sich, C., Grüne, M. et al. Sequential assignments in uniformly 13C- and 15N-labelled RNAs: The HC(N,P) and HC(N,P)-CCH-TOCSY experiments. J Biomol NMR 7, 251–255 (1996). https://doi.org/10.1007/BF00202042
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00202042