Hammouda Boualem, Worcester David
Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, MD, USA.
Biophys J. 2006 Sep 15;91(6):2237-42. doi: 10.1529/biophysj.106.083691. Epub 2006 Jun 30.
The helix-to-coil denaturation transition in DNA has been investigated in mixed solvents at high concentration using ultraviolet light absorption spectroscopy and small-angle neutron scattering. Two solvents have been used: water and ethylene glycol. The "melting" transition temperature was found to be 94 degrees C for 4% mass fraction DNA/d-water and 38 degrees C for 4% mass fraction DNA/d-ethylene glycol. The DNA melting transition temperature was found to vary linearly with the solvent fraction in the mixed solvents case. Deuterated solvents (d-water and d-ethylene glycol) were used to enhance the small-angle neutron scattering signal and 0.1M NaCl (or 0.0058 g/g mass fraction) salt concentration was added to screen charge interactions in all cases. DNA structural information was obtained by small-angle neutron scattering, including a correlation length characteristic of the inter-distance between the hydrogen-containing (desoxyribose sugar-amine base) groups. This correlation length was found to increase from 8.5 to 12.3 A across the melting transition. Ethylene glycol and water mixed solvents were found to mix randomly in the solvation region in the helix phase, but nonideal solvent mixing was found in the melted coil phase. In the coil phase, solvent mixtures are more effective solvating agents than either of the individual solvents. Once melted, DNA coils behave like swollen water-soluble synthetic polymer chains.
利用紫外光吸收光谱法和小角中子散射技术,在高浓度混合溶剂中研究了DNA的螺旋-线圈变性转变。使用了两种溶剂:水和乙二醇。发现对于质量分数为4%的DNA/d-水,“熔化”转变温度为94℃,对于质量分数为4%的DNA/d-乙二醇,“熔化”转变温度为38℃。在混合溶剂的情况下,发现DNA熔化转变温度随溶剂分数呈线性变化。使用氘代溶剂(d-水和d-乙二醇)来增强小角中子散射信号,并且在所有情况下都添加了0.1M NaCl(或质量分数0.0058 g/g)的盐浓度以屏蔽电荷相互作用。通过小角中子散射获得了DNA结构信息,包括含氢(脱氧核糖糖-胺基)基团之间的间距特征相关长度。发现该相关长度在熔化转变过程中从8.5增加到12.3 Å。发现乙二醇和水的混合溶剂在螺旋相的溶剂化区域中随机混合,但在熔化的线圈相中发现了非理想的溶剂混合。在线圈相中,溶剂混合物比任何一种单独的溶剂都是更有效的溶剂化剂。一旦熔化,DNA线圈的行为就像膨胀的水溶性合成聚合物链。