The metabolism of 17 beta-estradiol by lactoperoxidase: a possible source of oxidative stress in breast cancer.
Electron spin resonance (ESR) spectroscopy and oxygen consumption measurements using a Clark-type oxygen electrode have been used to study the metabolism of the estrogen 17 beta-estradiol by lactoperoxidase. Evidence for a one-electron oxidation of estradiol to its reactive phenoxyl radical intermediate is presented. The phenoxyl radical metabolite abstracts hydrogen from reduced glutathione generating the glutathione thiyl radical, which is spin trapped by 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and subsequently detected by ESR spectroscopy. In the absence of DMPO, molecular oxygen is consumed by a sequence of reactions initiated by the glutathione thiyl radical. Similarly, the estradiol phenoxyl radical abstracts hydrogen from reduced beta-nicotinamide-adenine dinucleotide (NADH) to generate the NAD. radical. The NAD. radical is not spin trapped by DMPO, but instead reduces molecular oxygen to the superoxide radical, which is then spin-trapped by DMPO. The superoxide generated may either spontaneously dismutate to form hydrogen peroxide or react with another NADH to form NAD., thus propagating a chain reaction leading to oxygen consumption and hydrogen peroxide accumulation. Ascorbate inhibits oxygen consumption when estradiol is metabolized in the presence of either glutathione or NADH by reducing radical intermediates back to their parent molecules and forming the relatively stable ascorbate radical. These results demonstrate that the futile metabolism of micromolar quantities of estradiol catalyzes the oxidation of much greater concentrations of biochemical reducing cofactors, such as glutathione and NADH, with hydrogen peroxide produced as a consequence. The accumulation of intracellular hydrogen peroxide could explain the hydroxyl radical-induced DNA base lesions recently reported for female breast cancer tissue.
Sipe
H J
HJ
Department of Chemistry, Hampden-Sydney College, Hampden-Sydney, VA 23943.
Jordan
S J
SJ
Hanna
P M
PM
Mason
R P
RP
D001205:Ascorbic Acid / Q000494:pharmacology; D001943:Breast Neoplasms / Q000378:metabolism*; D004958:Estradiol / Q000378:metabolism*; D005260:Female; D005609:Free Radicals; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007784:Lactoperoxidase / Q000502:physiology*; D009243:NAD / Q000494:pharmacology; D018384:Oxidative Stress; D013482:Superoxide Dismutase / Q000494:pharmacology
D005609:Free Radicals; D009243:NAD; D004958:Estradiol; D006861:Hydrogen Peroxide; D007784:Lactoperoxidase; D013482:Superoxide Dismutase; D005978:Glutathione; D001205:Ascorbic Acid