Sorokin Victor A, Valeev Vladimir A, Andrus Elena A, Gladchenko Galina O, Blagoi Yuri P
B. I. Verkin Institute for Low Temperature Physics and Engineering, National Academy of Sciences of Ukraine, 47 Lenin Ave., Kharkov 61103, Ukraine.
Macromol Biosci. 2004 May 17;4(5):532-8. doi: 10.1002/mabi.200400007.
Differential UV spectroscopy was used to study the temperature dependence of the conformational equilibrium in aqueous poly I . poly A . poly I (A2I) solutions containing Na+ (0.1-2 M) and Mg2+ (10(-5)-0.005 M) ions. Over the whole range of the studied Na+ and Mg2+ concentrations, the heating-induced destruction of the triple A2I helix is actually the A2I --> A + I + I (3 --> 1) transition. The rise of the transition temperature with increasing Na+ and Mg2+ contents is well described by Manning's and the "ligand" theories, which makes it possible to estimate the linear charge density on the single-stranded poly I (xi = 1.9 +/- 0.1) and the Mg2+-A2I binding constant (K = 1,250 M(-1) for the zero degree of binding). An analytical expression has been obtained, which correlates the constants of Mg2+ binding to three- and single-stranded polynucleotides (K3 and K1, respectively) and the linear charge density on them. There are only minor distinctions between the K3 values for A2U and A2I because these polynucleotides have similar structures. The difference in the K1 values is also slight as single-stranded poly U, poly I, and poly A have similar conformations. Dependence of the conformational transition temperatures of two triple helices with changing Mg2+ concentration.
采用差示紫外光谱法研究了含Na⁺(0.1 - 2 M)和Mg²⁺(10⁻⁵ - 0.005 M)离子的水性聚肌苷酸-聚腺苷酸-聚肌苷酸(A₂I)溶液中构象平衡的温度依赖性。在研究的Na⁺和Mg²⁺浓度的整个范围内,加热诱导的A₂I三螺旋破坏实际上是A₂I→A + I + I(3→1)转变。Manning理论和“配体”理论很好地描述了转变温度随Na⁺和Mg²⁺含量增加而升高的情况,这使得可以估计单链聚肌苷酸上的线性电荷密度(ξ = 1.9±0.1)以及Mg²⁺-A₂I结合常数(零结合度时K = 1250 M⁻¹)。得到了一个解析表达式,该表达式关联了Mg²⁺与三链和单链多核苷酸的结合常数(分别为K₃和K₁)以及它们上的线性电荷密度。A₂U和A₂I的K₃值之间只有微小差异因为这些多核苷酸具有相似的结构。K₁值的差异也很小,因为单链聚尿苷酸、聚肌苷酸和聚腺苷酸具有相似的构象。两种三螺旋的构象转变温度随Mg²⁺浓度变化的依赖性。