Wong Alan, Yan Zhimin, Huang Yining, Wu Gang
Department of Chemistry, Queen's University, 90 Bader lane, Kingston, Ontario, K7L 3N6, Canada.
Magn Reson Chem. 2008 Apr;46(4):308-15. doi: 10.1002/mrc.2136.
We report a solid-state (23)Na NMR study of monovalent cation (Li(+), Na(+), K(+), Rb(+), Cs(+) and NH(4) (+)) binding to double-stranded calf thymus DNA (CT DNA) at low relative humidity, ca 0-10%. Results from (23)Na--(31)P rotational echo double resonance (REDOR) NMR experiments firmly establish that, at low relative humidity, monovalent cations are directly bound to the phosphate group of CT DNA and are partially dehydrated. On the basis of solid-state (23)Na NMR titration experiments, we obtain quantitative thermodynamic parameters concerning the cation-binding affinity for the phosphate group of CT DNA. The free energy difference (DeltaG degrees ) between M(+) and Na(+) ions is as follows: Li(+) (-1.0 kcal mol(-1)), K(+) (7.2 kcal mol(-1)), NH(4) (+) (1.0 kcal mol(-1)), Rb(+) (4.5 kcal mol(-1)) and Cs(+) (1.5 kcal mol(-1)). These results suggest that, at low relative humidity, the binding affinity of monovalent cations for the phosphate group of CT DNA follows the order: Li(+) > Na(+) > NH(4) (+) > Cs(+) > Rb(+) > K(+). This sequence is drastically different from that observed for CT DNA in solution. This discrepancy is attributed to the different modes of cation binding in dry and wet states of DNA. In the wet state of DNA, cations are fully hydrated. Our results suggest that the free energy balance between direct cation-phosphate contact and dehydration interactions is important. The reported experimental results on relative ion-binding affinity for the DNA backbone may be used for testing theoretical treatment of cation-phosphate interactions in DNA.
我们报道了一项关于单价阳离子(Li⁺、Na⁺、K⁺、Rb⁺、Cs⁺和NH₄⁺)在低相对湿度(约0 - 10%)下与双链小牛胸腺DNA(CT DNA)结合的固态(²³)Na NMR研究。(²³)Na - (³¹)P旋转回波双共振(REDOR)NMR实验结果明确表明,在低相对湿度下,单价阳离子直接与CT DNA的磷酸基团结合且部分脱水。基于固态(²³)Na NMR滴定实验,我们获得了关于阳离子与CT DNA磷酸基团结合亲和力的定量热力学参数。M⁺和Na⁺离子之间的自由能差(ΔG°)如下:Li⁺(-1.0 kcal mol⁻¹)、K⁺(7.2 kcal mol⁻¹)、NH₄⁺(1.0 kcal mol⁻¹)、Rb⁺(4.5 kcal mol⁻¹)和Cs⁺(1.5 kcal mol⁻¹)。这些结果表明,在低相对湿度下,单价阳离子与CT DNA磷酸基团的结合亲和力顺序为:Li⁺ > Na⁺ > NH₄⁺ > Cs⁺ > Rb⁺ > K⁺。这个顺序与在溶液中观察到的CT DNA的顺序截然不同。这种差异归因于DNA干湿状态下阳离子结合模式的不同。在DNA的湿态中,阳离子是完全水合的。我们的结果表明,阳离子 - 磷酸基团直接接触和脱水相互作用之间的自由能平衡很重要。所报道的关于DNA主链相对离子结合亲和力的实验结果可用于检验DNA中阳离子 - 磷酸基团相互作用的理论处理。