Chemical determination of free radical-induced damage to DNA.
Free radical-induced damage to DNA in vivo can result in deleterious biological consequences such as the initiation and promotion of cancer. Chemical characterization and quantitation of such DNA damage is essential for an understanding of its biological consequences and cellular repair. Methodologies incorporating the technique of gas chromatography/mass spectrometry (GC/MS) have been developed in recent years for measurement of free radical-induced DNA damage. The use of GC/MS with selected-ion monitoring (SIM) facilitates unequivocal identification and quantitation of a large number of products of all four DNA bases produced in DNA by reactions with hydroxyl radical, hydrated electron, and H atom. Hydroxyl radical-induced DNA-protein cross-links in mammalian chromatin, and products of the sugar moiety in DNA are also unequivocally identified and quantitated. The sensitivity and selectivity of the GC/MS-SIM technique enables the measurement of DNA base products even in isolated mammalian chromatin without the necessity of first isolating DNA, and despite the presence of histones. Recent results reviewed in this article demonstrate the usefulness of the GC/MS technique for chemical determination of free radical-induced DNA damage in DNA as well as in mammalian chromatin under a vast variety of conditions of free radical production.
Dizdaroglu
M
M
Center for Chemical Technology, National Institute of Standards and Technology, Gaithersburg, MD 20899.
D000818:Animals; D003432:Cross-Linking Reagents / Q000378:metabolism; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000378:metabolism; D003855:Deoxyribose / Q000235:genetics*; D005609:Free Radicals; D008401:Gas Chromatography-Mass Spectrometry; D006878:Hydroxides; D017665:Hydroxyl Radical
D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D005609:Free Radicals; D006878:Hydroxides; D017665:Hydroxyl Radical; D003855:Deoxyribose