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Effect of Amino Acids on X-Ray-Induced Hydrogen Peroxide and Hydroxyl Radical Formation in Water and 8-Oxoguanine in DNA

I. N. Shtarkman1,2, S. V. Gudkov1, A. V. Chernikov1, and V. I. Bruskov1,2*

1Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia; fax: (496) 733-0553; E-mail: bruskov_vi@rambler.ru

2Pushchino State University, 142290 Pushchino, Moscow Region, Russia

* To whom correspondence should be addressed.

Received July 3, 2007; Revision received November 29, 2007
Generation of hydrogen peroxide and hydroxyl radicals in L-amino acid solutions in phosphate buffer, pH 7.4, under X-ray irradiation was determined by enhanced chemiluminescence in the luminol-p-iodophenol-peroxidase system and using the fluorescent probe coumarin-3-carboxylic acid, respectively. Amino acids are divided into three groups according to their effect on the hydrogen peroxide formation under irradiation: those decreasing yield of H2O2, having no effect, and increasing its yield. All studied amino acids at 1 mM concentration decrease the yield of hydroxyl radicals in solution under X-ray irradiation. However, the highest effect is observed in the order: Cys > His > Phe = Met = Trp > Tyr. At Cys, Tyr, and His concentrations close to physiological, the yield of hydroxyl radicals decreases significantly. Immunoenzyme analysis using monoclonal antibodies to 8-oxoguanine (8-oxo-7,8-dihydroguanine) was applied to study the effect of amino acids with the most pronounced antioxidant properties (Cys, Met, Tyr, Trp, Phe, His, Lys, Arg, Pro) on 8-oxoguanine formation in vitro under X-ray irradiation. It is shown that amino acids decrease the content of 8-oxoguanine in DNA. These amino acids within DNA-binding proteins may protect intracellular DNA against oxidative damage caused by formation of reactive oxygen species in conditions of moderate oxidative stress.
KEY WORDS: amino acids, X-ray irradiation, hydrogen peroxide, hydroxyl radicals, antioxidants, 8-oxoguanine in DNA

DOI: 10.1134/S0006297908040135