CRITERIA FOR THE MODE OF BINDING OF DNA-BINDING AGENTS
CRITERIA FOR THE MODE OF BINDING OF DNA-BINDING AGENTS
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DOI：
10.1016/0968-0896(95)00053-j
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发表时间：
1995-06-01
影响因子：
3.5
通讯作者：
CHAIRES, JB
中科院分区：
文献类型：
--
作者：
SUH, D;CHAIRES, JB
DNA结合剂的完整表征要求建立它们与DNA的结合模式。在缺乏高分辨率结构数据的情况下，结合模式通常是从各种溶液研究中间接推断出来的。这项研究的目的是证明只有某些方法才能可靠地推断DNA结合模式。描述了使用已证实的插层剂乙锭和沟槽粘结剂Hoechst 33258进行的比较荧光和流体动力学研究。我们的研究结果表明，虽然荧光强度、偏振和猝灭测量可以检测到配体与DNA的结合作用，但没有一个是结合模式的敏感指标。荧光接触能量转移研究可以可靠地指示插层作用，粘度测量也是如此。我们的结果表明，在缺乏高分辨率结构数据的情况下，可以使用可靠的标准来区分插层和沟槽结合。
A complete characterization of DNA binding agents requires that their mode of binding to DNA be established. In the absence of high resolution structural data, the mode of binding is, of necessity, usually inferred indirectly from various solution studies. The purpose of this study is to show that only certain methods can be used reliably to infer the DNA binding mode. Comparative fluorescence and hydrodynamic studies using the proven intercalator ethidium and the groove binder Hoechst 33258 are described. The results of our studies show that while fluorescence intensity, polarization, and quenching measurements can detect a binding interaction of the ligand with DNA, none are sensitive indicators of the binding mode. Fluorescence contact energy transfer studies can reliably indicate intercalation, as can viscosity measurements. Our results illustrate reliable criteria that may be used to distinguish intercalation from groove binding in the absence of high resolution structural data.