Glyoxal-guanine DNA adducts: detection, stability and formation in vivo from nitrosamines.
The glyoxal-deoxyguanosine adduct (gG) is formed from alpha-nitrosamino aldehydes and dG in vitro and in vivo from nitrosamines carrying the 2-hydroxyethyl side-chain as well as from N-nitrosomorpholine. The structures of all of the diastereomeric forms of both the cis and trans isomers of the adduct have been investigated by ab initio calculations and with nuclear magnetic resonance spectroscopy at 500 MHz. The preferred orientation of the OH groups is trans, but at equilibrium a small amount of the cis isomer was observed. The pH-independent equilibrium constant for the hydrolysis of the gG adduct is K = 1.36 x 10(-4) mol/L, and its rate of formation at pH 7.3 is k = 5.3 min-1 mol-1. In acid (pH 2), the hydrolysis of the nucleosidic linkage is nearly twice as rapid as the hydrolysis of gG to glyoxal and dG. We used a gG analogue to explore a number of reductive methods for derivatization of the adduct, but all of the processes either gave low yields or product mixtures which rendered them impractical for derivatizing the adduct in DNA. A 32P-postlabelling method for detection of the pH-sensitive gG adduct has been developed, which permitted detection of the adduct in the liver DNA of male Wistar rats after administration of selected nitrosamines. The levels of adducts found were: N-nitrosodiethanolamine > 2-hydroxyethylmethynitrosamine > N-nitrosomorpholine > 2-hydroxyethyethylnitrosamine. In separate experiments, N-nitrosodiethanolamine gave greater adduct levels than its metabolite 2-hydroxy-N-nitrosomorpholine. Mechanistic pathways for the generation of gG adducts in vitro and in vivo are discussed.
Loeppky
R N
RN
Department of Chemistry, University of Missouri, Columbia 65211, USA.
Cui
W
W
Goelzer
P
P
Park
M
M
Ye
Q
Q
D000818:Animals; D002273:Carcinogens / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid; D018736:DNA Adducts / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry; D003849:Deoxyguanosine / Q000737:chemistry / Q000378:metabolism*; D004052:Diethylnitrosamine / Q000031:analogs & derivatives / Q000633:toxicity; D007536:Isomerism; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008956:Models, Chemical; D008958:Models, Molecular; D009602:Nitrosamines / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D002273:Carcinogens; D018736:DNA Adducts; D009602:Nitrosamines; C014129:N-nitrosodiethanolamine; D004052:Diethylnitrosamine; D003849:Deoxyguanosine