利用傅里叶变换核磁共振对酵母苯丙氨酸转移核糖核酸的亚氨基和氨基质子进行实时溶剂交换研究。
Real-time solvent exchange studies of the imino and amino protons of yeast phenylalanine transfer RNA by Fourier transform NMR.
作者信息
Johnston P D, Figueroa N, Redfield A G
出版信息
Proc Natl Acad Sci U S A. 1979 Jul;76(7):3130-4. doi: 10.1073/pnas.76.7.3130.
Abstract
Real-time solvent exchange measurements using Fourier transform NMR at 270 MHz are presented. By means of the fast gel filtration column techniques originally developed for tritium exchange experiments, we were able to replace the solvent of a tRNA sample from an 1H2O to an 2H2O buffer and obtain a useful spectrum in 2-5 min. At 15 degrees C, there are 5 +/- 1 lowfield (-11 to -15 ppm relative to 2,2-dimethyl-2-silapentane-5-sulfonate) imino protons with exchange half times of minutes to hours. In addition, the m7G-46 C(8) proton and several amino protons are observed to exchange with similar rates. Analogous studies on unfractionated yeast tRNA suggest that such a class of slowly exchanging imino protons is present in several tRNAs, and that the activation energy for exchange is small [[approximatley 5 kcal/mol (21 kJ/mol)]. We speculate that these imino resonances arise from D-stem protons and that their slow exchange reflects stabilization by the numerous tertiary interactions involving this stem and the Mg2+ bound at the P-10 bend.
摘要
本文介绍了使用270兆赫傅里叶变换核磁共振进行实时溶剂交换测量的方法。借助最初为氚交换实验开发的快速凝胶过滤柱技术，我们能够将tRNA样品的溶剂从1H2O替换为2H2O缓冲液，并在2至5分钟内获得有用的光谱。在15摄氏度时，有5±1个低场（相对于2,2-二甲基-2-硅戊烷-5-磺酸盐为-11至-15 ppm）亚氨基质子，其交换半衰期为几分钟至几小时。此外，观察到m7G-46 C(8)质子和几个氨基质子以相似的速率进行交换。对未分级酵母tRNA的类似研究表明，几类tRNA中都存在这类缓慢交换的亚氨基质子，且交换的活化能较小[约5千卡/摩尔（21千焦/摩尔）]。我们推测这些亚氨基共振来自D-茎质子，其缓慢交换反映了涉及该茎和在P-10弯曲处结合的Mg2+的众多三级相互作用所导致的稳定性。
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Real-time solvent exchange studies of the imino and amino protons of yeast phenylalanine transfer RNA by Fourier transform NMR.利用傅里叶变换核磁共振对酵母苯丙氨酸转移核糖核酸的亚氨基和氨基质子进行实时溶剂交换研究。
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