Oxidation of bovine serum albumin initiated by the Fenton reaction--effect of EDTA, tert-butylhydroperoxide and tetrahydrofuran.
Oxidation of bovine serum albumin (BSA) was investigated using different oxidants: The water-soluble azo-initiator 2,2'azo-bis-(2-amidinopropane) hydrochloride (AAPH), a combination of FeCl(3) and ascorbate or the Fenton oxidant consisting of FeCl(2), H(2)O(2) and EDTA. In addition, the effects of exogenous compounds such as tert-butyl hydroperoxide (tBuOOH) or solvents such as tetrahydrofuran (THF), often used in model systems, was evaluated. The extent of protein damage was studied by measuring protein carbonyl groups and protein hydroperoxides. The interaction between Fenton oxidant and EDTA, THF or tBuOOH was further characterized using spin trapping electron spin resonance (ESR) spectroscopy. The results showed that the extent of protein oxidation depended on the oxidant used. The Fenton oxidant was the most reactive of the initiators tested. However, in the absence of EDTA, the Fenton system produced protein carbonyl groups on BSA equivalent to that obtained with the other oxidants, however, significantly more protein hydroperoxide was produced. Surprisingly, it was also found that addition of tBuOOH or THF to BSA reduced protein damage when the oxidation was initiated with the Fenton oxidant. ESR investigation showed that EDTA played a key role in the generation of free radicals. It was also revealed that in an EDTA containing system both tBuOOH and THF were able to react with radicals without inducing protein damage in effect protecting BSA from oxidative damage.
Baron
Caroline P
CP
Department of Seafood Research, Danish Institute for Fisheries Research, Technical University of Denmark, Building 221, Kgs, Lyngby DK-2800, Denmark. cba@dfu.min.dk
Refsgaard
Hanne H F
HH
Skibsted
Leif H
LH
Andersen
Mogens L
ML
D000818:Animals; D002417:Cattle; D004492:Edetic Acid / Q000494:pharmacology*; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals; D005663:Furans / Q000494:pharmacology*; D006861:Hydrogen Peroxide; D007501:Iron; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress / Q000502:physiology*; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000187:drug effects; D013113:Spin Labels; D020122:tert-Butylhydroperoxide / Q000494:pharmacology*
C045076:Fenton's reagent; D005609:Free Radicals; D005663:Furans; D013113:Spin Labels; D012710:Serum Albumin, Bovine; C018674:tetrahydrofuran; D020122:tert-Butylhydroperoxide; D004492:Edetic Acid; D006861:Hydrogen Peroxide; D007501:Iron