Double-strand hydrolysis of DNA by a magnesium(II) complex with diethylenetriamine

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• 作者：Pin Yang ; Rui Ren ; Maolin Guo ; Aixin Song ; Xiangli Meng ; Caixia Yuan ; Qinghua Zhou ; Huili Chen ; Zhenhai Xiong and Xiaoli Gao
• 关键词：Magnesium(II) ; Diethylenetriamine (dien) ; DNA ; Double ; strand ; Hydrolysis
• 刊名：Journal of Biological Inorganic Chemistry
• 出版年：2004
• 出版时间：June 2004
• 年：2004
• 卷：9
• 期：4
• 页码：495-506
• 全文大小：423 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Biochemistry
  Microbiology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-1327

文摘

The development of artificial nucleases that hydrolyze DNA or RNA is of great interest in molecular biology, biotechnology, and medicine. We now report that a magnesium(II) complex of diethylenetriamine (Mg-dien) can effectively promote the double-stranded cleavage of plasmid DNA and the dideoxynucleotide dApdA under physiological conditions of pH and temperature. Experiments performed in the presence of hydrogen peroxide, radical scavengers, or under rigorously anaerobic conditions indicate that DNA cleavage mediated by Mg-dien occurs via a hydrolytic path. Mg-dien efficiently hydrolyzes supercoiled pBR322 DNA and the pseudo-first-order rate constant at 37 °C and pH 8.0 is estimated to be 1.60 h–1. The dinucleotide dApdA hydrolysis, with Mg-dien at 170 M, shows a rate enhancement factor of ca. 5×108. 1H and 31P{1H} NMR studies show that Mg-dien effectively hydrolyzes 5-dAMP to give deoxyadenosine and inorganic phosphate. While Mg2+ has been found at the catalytic sites of many natural nucleases, Mg-dien appears to be the first synthetic Mg2+-containing system capable of hydrolyzing dideoxynucleotides and DNA and thus may provide a simple model system to assist mechanistic studies of naturally occurring nucleases.