Copper-phenanthroline-induced site-specific oxygen-radical damage to DNA. Detection of loosely bound trace copper in biological fluids

Previous studies have shown that the chelating agent 1,10-phenanthroline binds copper(II) ions and in the presence of a reducing agent and molecular °2 degrades DNA

J M C Gutteridge

2015

Scholarcy highlights

  • Previous studies have shown that the chelating agent 1,10-phenanthroline binds copper(II) ions and in the presence of a reducing agent and molecular °2 degrades DNA
  • The reaction mixture contained the following reagents added in the order stated: 0.4ml of calf thymus DNA, 0.1 ml of 1, 10-phenanthroline, 0.05 ml of NaN3, 0.05ml of copper salt or sample, 0.2ml of phosphate buffer, pH6.4, and O.1rml of mercaptoethanol
  • Thiobarbituric acid-reactive material released from DNA by copper-phenanthroline has fluorescent properties indistinguishable from those of the material formed between malondialdehyde and thiobarbituric acid
  • Previous studies have shown that H202 is an important intermediate in the degradation of DNA by 1,10phenanthroline and copper, since the reaction is substantially inhibited by catalase
  • Detection of trace copper in biological fluids should be included for each sample to detect other thiobarbituric acid-reactive materials
  • Normal human serum stored at - 20°C contained no loosely bound copper available to 1,10-phenanthroline, showing that 1,10-phenanthroline under these conditions does not remove copper from caeruloplasmin
  • Two samples of sweat obtained from athletes after severe exercise contained substantial concentrations of loosely bound copper

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