Wilson D H, Price H L, Henderson J, Hanlon S, Benight A S
Department of Chemistry, University of Illinois, Chicago 60680.
Biopolymers. 1990 Feb 5;29(2):357-76. doi: 10.1002/bip.360290208.
Dynamic and static light scattering, CD, and optical melting experiments have been conducted on M13mp19 viral circular single-strand DNA as a function of NaCl concentration. Over the 10,000-fold range in concentration from 100 microM to 1.0 M NaCl, the melting curves and CD spectra indicate an increase in base stacking and stability of stacked regions with increased salt concentration. Analysis of dynamic light scattering measurements of the single-strand DNA solutions as a function of K2 from 1.56 to 20 X 10(10) cm-2 indicates the collected autocorrelation functions are biexponential, thus revealing the presence of two decaying dynamic components. These components are taken to correspond to (1) global translational motions of the molecular center of mass and (2) motions of the internal molecular subunits. From the evaluated relaxation rates of these components, diffusion coefficients D0 and Dplat are determined. The center of mass translational diffusion coefficient D0, varies in a nonmonotonic manner, by 10%, from 3.75 X 10(-8) to 3.39 X 10(-8) cm2/s over the NaCl concentration range from 100 microM to 1.0 M. Likewise, the radius of gyration RG, obtained from static light scattering experiments, varies by 15% from 699 to 830 A over the same NaCl range Dplat, the diffusion coefficient of the internal subunits, displays a different dependence on the NaCl concentration and decreases, by nearly 22% in a titratable fashion, from 12.46 X 10(-8) to 10.26 X 10(-8) cm2/s, when the salt is increased from 100 microM to 1.0 M. A semiquantitative interpretation of these results is provided by analysis of the light scattering data in terms of the circular Rouse-Zimm chain. Rouse-Zimm model parameters are estimated from the experimental results, assuming the circular chains are composed of a fixed number of Gaussian segments, N + 1 = 15. The rms displacement of the internal segments, b, is estimated to be the smallest (442 A) in 100 mM NaCl. Increases of b to 467 A in 100 microM and 524 A in 1.0 M NaCl are observed. Meanwhile, the hypothetical friction factor of the internal subunits, f, progressively increases as the NaCl concentration is raised. It is inferred from the evaluated Rouse-Zimm model parameters that both the static flexibility of the circular chain and diffusive displacements of the internal subunits decrease with increases in NaCl concentration from 100 mM to 1.0 M.(ABSTRACT TRUNCATED AT 400 WORDS)
已针对M13mp19病毒环状单链DNA,在不同NaCl浓度下进行了动态光散射、静态光散射、圆二色(CD)和光学熔解实验。在NaCl浓度从100微摩尔至1.0摩尔的10000倍范围内,熔解曲线和CD光谱表明,随着盐浓度增加,碱基堆积以及堆积区域的稳定性增强。对单链DNA溶液在K2从1.56至20×10¹⁰ cm⁻²范围内的动态光散射测量分析表明,所收集的自相关函数是双指数的,从而揭示出存在两个衰减的动态成分。这些成分被认为分别对应于:(1)分子质心的整体平移运动;(2)分子内部亚基的运动。根据这些成分的评估弛豫速率,确定了扩散系数D0和Dplat。在100微摩尔至1.0摩尔的NaCl浓度范围内,质心平移扩散系数D0以非单调方式变化,变化幅度为10%,从3.75×10⁻⁸至3.39×10⁻⁸ cm²/s。同样,通过静态光散射实验获得的回转半径RG,在相同的NaCl浓度范围内,从699 Å变化至830 Å,变化幅度为15%。内部亚基的扩散系数Dplat对NaCl浓度呈现出不同的依赖性,当盐浓度从100微摩尔增加至1.0摩尔时,以可滴定的方式下降近22%,从12.46×10⁻⁸降至10.26×10⁻⁸ cm²/s。通过根据圆形Rouse-Zimm链对光散射数据进行分析,对这些结果进行了半定量解释。假设圆形链由固定数量(N + 1 = 15)的高斯链段组成,根据实验结果估算了Rouse-Zimm模型参数。内部链段的均方根位移b在100 mM NaCl中估计最小(442 Å)。在100微摩尔和1.0摩尔NaCl中,b分别增加至467 Å和524 Å。同时,随着NaCl浓度升高,内部亚基的假设摩擦因子f逐渐增大。根据评估的Rouse-Zimm模型参数推断,当NaCl浓度从100 mM增加至1.0摩尔时,圆形链的静态柔韧性和内部亚基的扩散位移均降低。(摘要截取自400字)