A simple copper(II)-L-histidine system for efficient hydrolytic cleavage of DNA.
Copper(II)-L-histidine complexes effectively promote the cleavage of plasmid DNA and dideoxynucleotide dApdA at physiological pH and temperature. Studies of the mechanism of plasmid DNA cleavage by added radical scavengers, using rigorously anaerobic experiments, analyses for malondialdehyde-like products, religation assays, and HPLC analyses, indicate that DNA cleavage mediated by Cu(L-His) occurs via a hydrolytic path. The hydrolytic cleavage rate constants at 37 degrees C are estimated to be 0.76 h-1 for the decrease of form I and 0.25 h-1 for the increase of form III. The phosphoimager picture reveals that Cu(L-His) cleaves DNA with a certain sequence specificity (preferentially at 5'-GT-3'). The dinucleotide hydrolysis shows, with [Cu(L-His)] = 0.8 mM, rate enhancement factors of > 10(8). Interestingly, histidine-metal ion interactions (with Cu(II), Ni(II), Zn(II), etc.) have been used for various applications, e.g., protein purification, cross-linking, and targeting proteins to lipid bilayers. Our findings may provide the basis for developing new applications and new ways to design more effective and useful catalysts for DNA cleavage. Cu(L-His) is one of only a few well-defined metal complexes demonstrated to hydrolytically cleave dideoxynucleotides and DNA.
Ren
R
R
Institute of Molecular Science, Shanxi University, Taiyuan 030006, People's Republic of China.
Yang
P
P
Zheng
W
W
Hua
Z
Z
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003300:Copper / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D009994:Osmolar Concentration; D010957:Plasmids; D013696:Temperature
D006639:Histidine; D003300:Copper; D004247:DNA