Frydman L, Rossomando P C, Frydman V, Fernandez C O, Frydman B, Samejima K
Facultad de Farmacia y Bioquimica, Universidad de Buenos Aires, Argentina.
Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9186-90. doi: 10.1073/pnas.89.19.9186.
15N NMR spectroscopy was used to explore the interactions between natural polyamines and Escherichia coli tRNA. It was found that when tRNA is added to solutions of 15N-labeled spermine or spermidine, there is a considerable decrease in the relative heights of the -NH(2+)--resonances with respect to the signals arising from the -NH3+ groups. The presence of tRNA was also found to reduce the longitudinal relaxation times T1 of the nitrogens, mainly those of the -NH(2+)- groups. The longitudinal relaxation times of the nitrogens were used to characterize the temperature dependence of the binding, and they allowed us to calculate the activation energies that determine the correlation times of amino groups in the presence of tRNA. Both the thermodynamic and the relaxation results indicate that (i) spermine binds more strongly to tRNA than spermidine does and (ii) within each of these molecules the -NH(2+)- groups bind more strongly to tRNA than the more electropositive -NH3+ moieties. This specificity suggests that the interaction between polyamines and tRNA cannot be described exclusively in terms of electrostatic forces and that other interactions (most likely, hydrogen bonding) are very important for establishing the polyamine-tRNA link. Some of the factors that may conspire against the binding of -NH3+ groups to tRNA are briefly discussed.
利用15N核磁共振光谱法探究天然多胺与大肠杆菌转运核糖核酸(tRNA)之间的相互作用。研究发现,当将tRNA添加到15N标记的精胺或亚精胺溶液中时,相对于 -NH3+ 基团产生的信号,-NH(2+)--共振的相对高度会显著降低。还发现tRNA的存在会降低氮原子的纵向弛豫时间T1,主要是 -NH(2+)- 基团的纵向弛豫时间。利用氮原子的纵向弛豫时间来表征结合的温度依赖性,并据此计算出在tRNA存在下决定氨基相关时间的活化能。热力学和弛豫结果均表明:(i)精胺与tRNA的结合比亚精胺更强;(ii)在这些分子中,-NH(2+)- 基团与tRNA的结合比电正性更强的 -NH3+ 部分更强。这种特异性表明,多胺与tRNA之间的相互作用不能仅用静电力来描述,其他相互作用(很可能是氢键)对于建立多胺 - tRNA连接非常重要。文中简要讨论了一些可能不利于 -NH3+ 基团与tRNA结合的因素。