期刊
JOURNAL OF BIOMOLECULAR NMR
卷 39, 期 1, 页码 17-29出版社
SPRINGER
DOI: 10.1007/s10858-007-9167-5
关键词
NMR spectroscopy; isotope labeled RNA; chi-angle determination; cross-correlated relaxation
A novel NMR pulse sequence is introduced to determine the glycosidic torsion angle v in C-13, N-15-labeled oligonucleotides. The quantitative Gamma-HCNCH measures the dipolar cross-correlated relaxation rates Gamma(DD,DD)(C8H8,C1'H1') (pyrimidines) and Gamma(DD,DD)(C8H8,C1'H1') (purines). Cross-correlated relaxation rates of a C-13, N-15-labeled RNA 14mer containing a cUUCGg tetraloop were determined and yielded chi-angles that agreed remarkably well with data derived from the X-ray structure of the tetraloop. In addition, the method was applied to the larger stemloop D (SLD) subdomain of the Coxsackievirus B3 cloverleaf 30mer RNA and the effect of anisotropic rotational motion was examined for this molecule. It could be shown that the chi-angle determination especially for nucleotides in the anti conformation was very accurate and the method was ideally suited to distinguish between the syn and the anti-conformation of all four types of nucleotides.
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