ISOLATION OF A MALONDIALDEHYDE-DEOXYGUANOSINE ADDUCT FROM RAT-LIVER DNA

被引:26
作者
AGARWAL, S [1 ]
DRAPER, HH [1 ]
机构
[1] UNIV GUELPH,COLL BIOL SCI,DEPT NUTR SCI,GUELPH N1G 2W1,ONTARIO,CANADA
基金
加拿大自然科学与工程研究理事会;
关键词
LIPID PEROXIDATION; MALONDIALDEHYDE; LIVER DNA; DEOXYGUANOSINE; FREE RADICALS;
D O I
10.1016/0891-5849(92)90043-G
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In previous studies, an adduct of malondialdehyde (MDA) with guanine was identified in rat and human urine. Subsequent detection of an adduct with deoxyguanosine (dG) in urine prompted an investigation of its possible occurrence in DNA. Rat liver DNA was hydrolyzed using nuclease P1 and alkaline P-ase and subjected to deoxyribonucleoside analysis using reverse phase high-pressure liquid chromatography (HPLC) with fluorescence detection. A compound was isolated that could not be separated from a synthetic pyrimidinopurine adduct of MDA and dG (dG-MDA). Partial hydrolysis released guanine (Gua), Gua-MDA, and dG in amounts that, in aggregate, were the molar equivalent of the starting material calculated by fluorescence analysis as dG-MDA. Complete acid hydrolysis of the isolate yielded an equimolar amount of MDA. Analysis of liver DNA isolated from growing rats yielded a value for dG-MDA content of 9.0 +/- 1.6 pmol/100 mug DNA (xBAR +/- SEM, N = 5). This value is approximately 7 times those reported for the 8-hydroxy deoxyguanosine content of rat liver nuclear DNA. This study demonstrates that DNA is modified in vivo by reactions of its guanylate moiety with MDA, and indicates that, at least in the case of rat liver DNA, the prevalence of such modifications is greater than those caused by reactions with hydroxyl radicals.
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页码:695 / 699
页数:5
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