Selective detection of 2-deoxyribonolactone in DNA.
2-Deoxyribonolactone (L) is an oxidized abasic lesion that is produced by a variety of DNA damaging agents. It exhibits unique biological effects with respect to its proclivity to form DNA-protein cross-links and promutagenic base pairs. Recent evidence suggests that the levels of this lesion caused by oxidative stress are underestimated. We have developed a simple, selective method for detecting subpicomole amounts of L in DNA. The method takes advantage of the selective reaction of the butenolide (2) derived from beta-elimination from L with a biotinylated derivative of cysteine. This method will be useful for analyzing the levels of this oxidized abasic site in DNA.
Sato
Kousuke
K
Department of Chemistry, Johns Hopkins University, 3400 North Charles Street, Baltimore, Maryland 21218, USA.
Greenberg
Marc M
MM
D001345:Autoradiography; D001710:Biotin / Q000031:analogs & derivatives; D003545:Cysteine / Q000031:analogs & derivatives; D004247:DNA / Q000737:chemistry*; D004249:DNA Damage; D008401:Gas Chromatography-Mass Spectrometry; D006735:Horseradish Peroxidase / Q000737:chemistry; D010084:Oxidation-Reduction; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013400:Sugar Acids / Q000032:analysis*
D013400:Sugar Acids; C056405:2,4,5-trihydroxypentanoic acid gamma-lactone; D001710:Biotin; D004247:DNA; D006735:Horseradish Peroxidase; D003545:Cysteine