Experiments for Correlating Quaternary Carbons in RNA Bases

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Authors

FIALA Radovan MUNZAROVÁ Markéta SKLENÁŘ Vladimír

Year of publication 2004
Type Article in Periodical
Magazine / Source Journal of Biomolecular NMR
MU Faculty or unit

Faculty of Science

Citation
Field Biochemistry
Keywords ab initio; correlation; DFT; NMR; quternary carbon; RNA; spin-spin coupling
Description The paper presents a set of triple-resonance two-dimensional experiments 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 (the 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 performance of the proposed approach is demonstrated on a 0.5 mM 25-mer RNA. The results show that the experiments are applicable to samples containing agents for weak molecular alignment. The design of the correlation experiments has been supported by ab initio calculations of scalar spin-spin couplings for the AU and GC base pairs. The ab initio data reveal surprisingly high values of guanine 2JN1C5 and uracil 2JN3C5 couplings that are in a qualitative agreement with the experimental data. The sensitivity of the spin-spin couplings to base pairing as well as the agreement with the experiment depend strongly on the type of nuclei involved and the number of bonds separating them.
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