NUCLEASE ACTIVITY OF 1,10-PHENANTHROLINE-COPPER ION

1，10-菲咯啉-亚铜络合物，（OP）2Cu+，与过氧化氢作为 共反应物，是一种人工核酸酶，其通过一种酶切割DNA和RNA。 氧化途径在生理pH值和温度。 以来 反应通过核酸-（OP）2Cu+络合物进行，核酸酶被 这取决于核酸的三维结构。 为 例如，A-DNA的切割速率是B-DNA的1/7;单链和 Z-DNA对切割有抵抗力。 由于（OP）2Cu+很小， 结构紧凑，其核酸酶活性应受局部 DNA构象，而不是与切割远端碱基的任何相互作用 绝佳的价钱 因此，（OP）2Cu+可以提供一种独特的方法来映射局部 DNA结构的变化，可能是功能上重要的， 基因信息的表达。 这项赠款的第一个目标是确定结构性决定因素 它通过研究DNA的消化模式来控制DNA的断裂 已知三维结构的寡核苷酸。 然后是乳沟 不同细菌操纵子之间的关系将决定 用（OP）2Cu+检测。 在这些分子中引入的构象变化 将通过突变和配体结合来分析区域。 的范围 人工核酸酶活性将通过连接 1，10-菲咯啉与DNA的特异性配体结合，并将这些配体 载体转化为位点特异性核酸酶。 例如，OP，绑定到 针对Z-DNA的抗体，应该特异性地在 左手结构 最后，细胞内作用的效率 这种人工核酸酶将探索与检查的目标 DNA原位结构和设计细胞类型特异性细胞毒性剂。

1,10-Phenanthroline-cuprous complex, (OP)2Cu+, with hydrogen peroxide as a coreactant, is an artificial nuclease which cleaves DNA and RNA by an oxidative pathway at physiological pH's and temperatures. Since the reaction proceeds via a nucleic acid-(OP)2Cu+ complex, the nuclease is dependent on the three-dimensional structure of the nucleic acid. For example, A-DNA is cleaved at 1/7th the rate of B-DNA; single stranded and Z-DNA's are resistant to cleavage. Since (OP)2Cu+ is small and structurally compact, its nuclease activity should be governed by the local DNA conformation and not any interaction with bases distal to the cleavage site. Therefore, (OP)2Cu+ may provide a unique method to map the local variations in DNA structure that may be functionally important in the expression of genetic information. The first goal of this grant is to identify the structural determinants which control DNA scission by studying the digestion patterns of oligonucleotides of known three dimensional structure. Then, the cleavage of different bacterial operons will determine if common structural features are detected by (OP)2Cu+. Conformational changes introduced in these regions by mutations and ligand binding will be analyzed. The scope of the artificial nuclease activity will be enhanced by attaching 1,10-phenanthroline to specific ligands of DNA and converting these carriers into site specific nucleases. For example, OP, bound to antibodies directed against Z-DNA, should specifically nick this left-handed structure. Finally, the efficiency of the intracellular action of this artificial nuclease will be explored with the goals of examining DNA structure in situ and designing cell-type specific cytotoxic agents.