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肽核酸(PNA)/DNA三链螺旋的熔解热力学

Thermodynamics of the melting of PNA(2)/DNA triple helices.

作者信息

Krupnik O V, Guscho Y, Sluchanko K, Nielsen P, Lazurkin Y

机构信息

Institute of Molecular Genetics, Russian Academy of Sciences, Kurchatov sq. 2, Moscow 123182, Russia.

出版信息

J Biomol Struct Dyn. 2001 Dec;19(3):535-42. doi: 10.1080/07391102.2001.10506761.

Abstract

Equilibrium melting curves were obtained for triplexes, formed by single stranded DNA containing an A10 target with bis-PNA consisting of two T10 decamers. Thermodynamic parameters of melting were determined for Na(+) concentrations 50, 200 and 600mM by two methods. The melting enthalpy Delta H degrees was evaluated from the width of the differential melting curves and from the concentration dependence of the melting temperature. The latter method allowed also evaluating the melting entropy Delta S degrees. The following results were obtained: Delta H degrees = - 137 kcal/M, Delta S degrees = - 368 cal/M.K, Delta G degrees (298)= - 27 kcal/M. No dependence of Delta H degrees, Delta S degrees and Delta G degrees (298) was observed upon ionic strength within the accuracy of the experiment (+/- 10%). The absolute values of Delta H degrees, Delta S degrees and Delta G degrees(298) are 2 to 3 times higher than the published values of corresponding melting parameters for decameric PNA/DNA duplexes of various nucleic base sequences. The origin of the extremely high stability of the triplexes is discussed.

摘要

获得了由包含A10靶标的单链DNA与由两个T10十聚体组成的双PNA形成的三链体的平衡熔解曲线。通过两种方法测定了50、200和600mM Na(+)浓度下的熔解热力学参数。从微分熔解曲线的宽度和熔解温度的浓度依赖性评估熔解焓ΔH°。后一种方法还可以评估熔解熵ΔS°。得到以下结果:ΔH° = - 137 kcal/M,ΔS° = - 368 cal/M·K,ΔG°(298) = - 27 kcal/M。在实验精度(±10%)范围内,未观察到ΔH°、ΔS°和ΔG°(298)对离子强度的依赖性。ΔH°、ΔS°和ΔG°(298)的绝对值比各种核酸碱基序列的十聚体PNA/DNA双链体相应熔解参数的已发表值高2至3倍。讨论了三链体极高稳定性的起源。

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