β-carotene and protein oxidation:: effects of ascorbic acid and α-tocopherol

The effect of beta-carotene on protein oxidation was examined under different oxygen (O-2) tensions and with other antioxidants: alpha-tocopherol, ascorbic acid, and mixtures of antioxidants. Human serum albumin (HSA) was incubated with 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) to induce protein oxidation (carbonyl formation), under 15, 150, and 760 torr of O-2 tension. Antioxidant activity was related to O-2 tension, antioxidant concentrations and interaction between mixtures of antioxidants: (1) Under 15 torr of O-2, incubating HSA with AAPH, 1.6 mu M beta-carotene, 80 mu M alpha-tocopherol, 160 mu M ascorbic acid, and mixtures (0.1 mu M beta-carotene, 5.0 CIM alpha-tocopherol and 10 mu M ascorbic acid) resulted in 24, 29, 39, and 44% reduction of carbonyl formation, respectively. (2) Under 150 torr of O-2 tension, the antioxidant effect of beta-carotene was decreased by 4% but increasing O-2 tension did not diminish the antioxidant effects of alpha-tocopherol, ascorbic acid, or antioxidant mixtures. (3). Under 760 torr of O-2 tension, adding 1.6 mu M beta-carotene resulted in 26% more carbonyl formation. (4) Under 760 torr of O-2 tension, the antioxidant effect of ascorbic acid was decreased 32% compared to what was observed at 150 torr of O-2 tension. Changes in O-2 tension had no effect on the antioxidant effect of alpha-tocopherol. The mixture of antioxidants inhibited carbonyl formation by 37% and was 7% less effective than that of 15 and 150 torr of O-2 tension. High concentration of beta-carotene produces more protein oxidation in the presence of high O-2 tension by a prooxidant mechanism. Mixtures of beta-carorene, alpha-tocopherol, and ascorbic acid provided better protective effects on protein oxidation than any single compound. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.