Mascotti D P, Lohman T M
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Biochemistry. 1997 Jun 10;36(23):7272-9. doi: 10.1021/bi970272n.
We have examined the equilibrium binding of a series of synthetic oligoarginines (net charge z = +2 to +6) containing tryptophan to poly(U), poly(A), poly(C), poly(I), and double-stranded (ds) DNA. Equilibrium association constants, K(obs), measured by monitoring tryptophan fluorescence quenching, were examined as functions of monovalent salt (MX) concentration and type, as well as temperature, from which deltaG(standard)obs, deltaH(obs), and deltaS(standard)obs were determined. For each peptide, K(obs) decreases with increasing [K+], and the magnitude of the dependence of K(obs) on [K+], delta log K(obs)/delta log[K+], increases with increasing net peptide charge. In fact, the values of delta log K(obs)/delta log[K+] are equivalent for oligolysines and oligoarginines possessing the same net positive charge. However, the values of K(obs) are systematically greater for oligoarginines binding to all polynucleotides, when compared to oligolysines with the same net charge. The origin of this difference is entirely enthalpic, with deltaH(obs), determined from van't Hoff analysis, being more exothermic for oligoarginine binding. The values of deltaH(obs) are also independent of [K+]; therefore, the salt concentration dependence of deltaG(standard)obs is entirely entropic in origin, reflecting the release of cations from the nucleic acid upon complex formation. These results suggest that hydrogen bonding of arginine to the phosphate backbone of the nucleic acids contributes to the increased stability of these complexes.
我们研究了一系列含色氨酸的合成寡聚精氨酸(净电荷z = +2至+6)与聚(U)、聚(A)、聚(C)、聚(I)和双链(ds)DNA的平衡结合。通过监测色氨酸荧光猝灭测量的平衡缔合常数K(obs),作为单价盐(MX)浓度和类型以及温度的函数进行了研究,由此确定了ΔG(标准)obs、ΔH(obs)和ΔS(标准)obs。对于每种肽,K(obs)随[K+]增加而降低,并且K(obs)对[K+]的依赖程度,即Δlog K(obs)/Δlog[K+],随肽净电荷增加而增大。事实上,具有相同净正电荷的寡聚赖氨酸和寡聚精氨酸的Δlog K(obs)/Δlog[K+]值是相等的。然而,与具有相同净电荷的寡聚赖氨酸相比,寡聚精氨酸与所有多核苷酸结合时的K(obs)值系统性地更高。这种差异的根源完全是焓性的,通过范特霍夫分析确定的ΔH(obs),对于寡聚精氨酸结合来说放热更多。ΔH(obs)的值也与[K+]无关;因此,ΔG(标准)obs对盐浓度的依赖性完全源于熵,反映了复合物形成时阳离子从核酸中的释放。这些结果表明精氨酸与核酸磷酸主链的氢键作用有助于这些复合物稳定性的提高。