Wilson W D, Wang Y H, Krishnamoorthy C R, Smith J C
Biochemistry. 1985 Jul 16;24(15):3991-9. doi: 10.1021/bi00336a029.
The binding of propidium to poly(dA).poly(dT) [poly(dA.dT)] and to poly[d(A-T)].poly[d(A-T)] [poly[d(A-T)2]] has been compared under a variety of solution conditions by viscometric titrations, binding studies, and kinetic experiments. The binding of propidium to poly[d(A-T)2] is quite similar to its binding to calf thymus deoxyribonucleic acid (DNA). The interaction with poly(dA.dT), however, is quite unusual. The viscosity of a poly(dA.dT) solution first decreases and then increases in a titration with propidium at 18 degrees C. The viscosity of poly[d(A-T)2] shows no decrease in a similar titration. Scatchard plots for the interaction of propidium with poly(dA.dT) show the classical upward curvature for positive cooperativity. The curvature decreases as the temperature is increased in binding experiments. A van't Hoff plot of the observed binding constants yields an apparent positive enthalpy of approximately +6 kcal/mol for the propidium-poly(dA.dT) interaction. Propidium binding to poly[d(A-T)2] shows no evidence for positive cooperativity, and the enthalpy change for the reaction is approximately -9 kcal/mol. Both the magnitude of the dissociation constants and the effects of ionic strength are quite similar for the dissociation of propidium from poly(dA-T)2] and from poly[d(A-T)2], suggesting that the intercalated states are similar for the two complexes. The observed association reactions, under pseudo-first-order conditions, are quite different. Plots of the observed pseudo-first-order association rate constant vs. polymer concentration have much larger slopes for propidium binding to poly[d(A-T)2] than to poly(dA.dT).(ABSTRACT TRUNCATED AT 250 WORDS)
在多种溶液条件下,通过粘度滴定、结合研究和动力学实验,比较了碘化丙啶与聚(dA)·聚(dT)[聚(dA·dT)]以及与聚[d(A - T)]·聚[d(A - T)][聚[d(A - T)₂]]的结合情况。碘化丙啶与聚[d(A - T)₂]的结合与其与小牛胸腺脱氧核糖核酸(DNA)的结合非常相似。然而,它与聚(dA·dT)的相互作用却很不寻常。在18℃下用碘化丙啶滴定聚(dA·dT)溶液时,其粘度先降低后升高。在类似滴定中,聚[d(A - T)₂]的粘度没有降低。碘化丙啶与聚(dA·dT)相互作用的Scatchard图显示出正协同作用的典型向上弯曲。在结合实验中,随着温度升高,曲率减小。观察到的结合常数的范特霍夫图显示,碘化丙啶 - 聚(dA·dT)相互作用的表观正焓约为 +6千卡/摩尔。碘化丙啶与聚[d(A - T)₂]的结合没有正协同作用的证据,该反应的焓变约为 -9千卡/摩尔。碘化丙啶从聚(dA - T)₂]和聚[d(A - T)₂]解离的解离常数大小以及离子强度的影响非常相似,这表明两种配合物的插入状态相似。在伪一级条件下观察到的缔合反应却大不相同。观察到的伪一级缔合速率常数与聚合物浓度的关系图显示,碘化丙啶与聚[d(A - T)₂]结合的斜率比与聚(dA·dT)结合的斜率大得多。(摘要截短于250字)