Rinnenthal Jörg, Richter Christian, Ferner Jan, Duchardt Elke, Schwalbe Harald
Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe-University, Max-von-Laue-Strasse 7, 60438 Frankfurt/Main, Germany.
J Biomol NMR. 2007 Sep;39(1):17-29. doi: 10.1007/s10858-007-9167-5. Epub 2007 Jul 20.
A novel NMR pulse sequence is introduced to determine the glycosidic torsion angle chi in (13)C,(15)N-labeled oligonucleotides. The quantitative Gamma-HCNCH measures the dipolar cross-correlated relaxation rates Gamma(DD,DD)(C8H8,C1'H1') (pyrimidines) and Gamma(DD,DD)(C6H6,C1'H1') (purines). Cross-correlated relaxation rates of a (13)C,(15)N-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.
引入了一种新型核磁共振脉冲序列来确定（13）C、（15）N标记的寡核苷酸中的糖苷扭转角χ。定量的Gamma-HCNCH测量偶极交叉相关弛豫率Gamma(DD,DD)(C8H8,C1'H1')（嘧啶）和Gamma(DD,DD)(C6H6,C1'H1')（嘌呤）。测定了含有cUUCGg四环的（13）C、（15）N标记的RNA 14聚体的交叉相关弛豫率，得到的χ角与从四环的X射线结构获得的数据非常吻合。此外，该方法应用于柯萨奇病毒B3苜蓿叶形30聚体RNA的较大茎环D（SLD）亚结构域，并研究了该分子的各向异性旋转运动的影响。结果表明，χ角的测定，特别是对于反式构象的核苷酸，非常准确，该方法非常适合区分所有四种类型核苷酸的顺式和反式构象。
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