Experiments for Correlating Quaternary Carbons in RNA Bases
| Authors | |
|---|---|
| Year of publication | 2004 |
| Type | Article in Proceedings |
| Conference | 19th NMR Valtice |
| MU Faculty or unit | |
| Citation | |
| Web | http://www.chemi.muni.cz/nmr/radek/nmrvaltice/nmrvaltice.htm |
| Field | Biochemistry |
| Keywords | NMR; spin-spin coupling; RNA; nucleic acid; quaternary carbon |
| Description | A set of triple-resonance two-dimensional NMR experiments is presented that can be used for correlating all quaternary carbons in RNA bases to one or more of the base protons. The experiments make use of either three-bond proton-carbon couplings and one selective INEPT step (long-range selective HSQC experiment) to transfer the magnetization between a proton and the carbon of interest and back, or they rely on one- and/or two-bond heteronuclear (the H(CN)C and H(N)C experiments) or carbon-carbon couplings (the H(C)C experiment) and multiple INEPT transfer steps. The effect of the large one-bond carbon-carbon coupling in t1 is removed by a constant time evolution or by a selective refocusing. The sensitivity of all the experiments proposed is high enough to obtain a spectrum of 0.5 mM 25-mer RNA stem-loop in four hours or less on a 600 MHz (1H) spectrometer. The results show that the experiments are also applicable to samples containing agents for weak molecular alignment. |
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