Brady G W, Satkowski M, Foos D, Benham C J
Center for Laboratories & Research, New York State Department of Health, Albany 12201.
J Mol Biol. 1987 May 5;195(1):185-91. doi: 10.1016/0022-2836(87)90335-4.
The techniques of small-angle X-ray scattering and analysis that have been developed by the authors are used to investigate the influence of ionic strength on the superhelical conformation of native COP608 plasmid DNA in solution. For salt concentrations below 0.1 M, the superhelicity is partitioned between twisting (Tw) and writhing (Wr) in the ratio delta Tw/Wr = 2. Near the physiological salt concentration, [Na+] = 0.2 M, a co-operative transition is observed in which the pitch angle of the toroidal superhelix is drastically decreased. This results in an almost complete relaxation of writhe. At salt concentrations in excess of the threshold for this transition, the superhelical partitioning occurs in the ratio delta Tw/Wr greater than 25. Energetic considerations support the suggestion that this transition results from co-operative, superhelical B to Z transconformation reactions at susceptible sites. A method is discussed that will enable the direct measurement of this secondary structural transition by means of X-ray scattering.
作者开发的小角X射线散射和分析技术用于研究离子强度对溶液中天然COP608质粒DNA超螺旋构象的影响。对于盐浓度低于0.1M的情况,超螺旋度在扭曲(Tw)和缠绕(Wr)之间以δTw/Wr = 2的比例分配。在生理盐浓度[Na+]=0.2M附近,观察到一个协同转变,其中环形超螺旋的螺距角急剧减小。这导致缠绕几乎完全松弛。在超过该转变阈值的盐浓度下,超螺旋分配以δTw/Wr大于25的比例发生。能量考虑支持这样的观点,即这种转变是由易感位点处的协同超螺旋B到Z构象转变反应引起的。讨论了一种通过X射线散射直接测量这种二级结构转变的方法。