Oxidation reactions of a bovine serum albumin-bilirubin complex. A pulse radiolysis study.

Using the technique of pulse radiolysis, oxidation studies of the bovine serum albumin-bilirubin (BSA-BR) system with radicals like CCl3OO., N3., (SCN)2.-, Br2.- and OH. generated in neutral and alkaline medium are reported. In a neutral solution, BSA protects the bound BR very efficiently from the attack of these radicals. The experimental k/k' values for the reaction of CCl3OO., N3. and Br2.- radicals are 2.46, 1.78 and 2.55 respectively, where k and k' are the bimolecular rate constants for the formation of the semi-oxidized BSA and BR radicals respectively. The calculated ratios from our measurements of rate constants k and k' are 0.16, 0.28 and 1.38 for CCl3OO., N3. and Br2.- respectively. These values indicate protection of BR by BSA from free radical attack. For Br2.- radical-induced oxidation of the BSA-BR system, a radical transfer from protein to BR was observed. OH. shows very fast adduct formation with both BSA and BR. The bimolecular rate constant for the formation of BR-OH adducts at PH 8+/- 0.2 is 9.5 x 10(9) dm3 mol-1 s-1 (540 nm). OH. adds to BSA at neutral pH with a rate constant of 3.0 +/- 1.0 x 10(10) dm3 mol-1 s-1 (305 nm). In the BSA-BR complex, BSA fully protects BR from OH. attack and the (BSA-BR)-OH adduct further reacts with free BR molecule if present in solution.