Determination of sugar conformation in large RNA oligonucleotides from analysis of dipole-dipole cross correlated relaxation by solution NMR spectroscopy

被引:25
|
作者
Richter, C
Griesinger, C
Felli, I
Cole, PT
Varani, G
Schwalbe, H
机构
[1] MIT, Dept Chem, Francis Bitter Natl Magnet Lab, Cambridge, MA 02139 USA
[2] MRC, Mol Biol Lab, Cambridge CB2 2QH, England
[3] Goethe Univ Frankfurt, Inst Organ Chem, D-60439 Frankfurt, Germany
关键词
anisotropic tumbling; cross correlated relaxation; isotope labelling; ribose ring puckering; RNA;
D O I
10.1023/A:1008319130714
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A new experiment, the forward directed quantitative Gamma-HCCH-TOCSY for the measurement of the conformation of the five-membered ribosyl unit in RNA oligonucleotides, is presented. The experiment relies on quantification of cross peak intensities caused by evolution of CH,CH-dipole-dipole cross correlated relaxation in non-evolution periods and the resolution enhancement obtainable in forward directed HCC-TOCSY transfer. Cross correlated relaxation rates are interpreted to reveal the sugar conformation of 22 out of 25 nucleotides in an isotopically labelled 25-mer RNA. The results obtained with this new method are in agreement with the conformational analysis derived from (3)J(H,H) coupling constants.
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页码:241 / 250
页数:10
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