Hydroxyl free radical adduct of deoxyguanosine: sensitive detection and mechanisms of formation.
DNA or 2-deoxyguanosine reacts with hydroxyl free radical to form 8-hydroxy-deoxyguanosine (8-OH-dG). We found that 8-OH-dG can be effectively separated from deoxyguanosine by high pressure liquid chromatography and very sensitively detected using electrochemical detection. The sensitivity of electrochemical detection is about one-thousand fold enhanced over optical detection. Utilizing deoxyguanosine in bicarbonate buffer it was found that ferrous ion, but not ferric ion, was effective in forming 8-OH-dG. The hydroxyl free radical scavenging agents, thiourea and ethanol, were very effective in quenching Fe(11) mediated 8-OH-dG formation, but superoxide dismutase had very little effect.
Floyd
R A
RA
Oklahoma Medical Research Foundation, Biomembrane Research Program, Oklahoma City 73104.
Watson
J J
JJ
Wong
P K
PK
Altmiller
D H
DH
Rickard
R C
RC
D000080242:8-Hydroxy-2'-Deoxyguanosine; D002851:Chromatography, High Pressure Liquid; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism*; D004563:Electrochemistry; D000431:Ethanol / Q000494:pharmacology; D016166:Free Radical Scavengers; D005609:Free Radicals; D017665:Hydroxyl Radical / Q000032:analysis / Q000378:metabolism*; D013890:Thiourea / Q000494:pharmacology
D016166:Free Radical Scavengers; D005609:Free Radicals; D017665:Hydroxyl Radical; D000431:Ethanol; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine; D013890:Thiourea