title: Protein radicals in the reaction between H2O2-activated metmyoglobin and bovine serum albumin.
abstract: Hydrogen peroxide activation of MMb with and without the presence of BSA gave rise to rapid formation of hyper-valent myoglobin species, myoglobin ferryl radical (*MbFe(IV) = O) and/or ferrylmyoglobin (MbFe(IV) = O). Reduction of MbFe(IV) = O showed first-order kinetics for a 1-2 times stoichiometric excess of H2O2 to MMb while a 3-10 times stoichiometric excess of H2O2 resulted in a biphasic reaction pattern. Radical species formed in the reaction between MMb, H2O2 and BSA were influenced by [H2O2] as measured by electron spin resonance (ESR) spectroscopy and resulted in the formation of cross-linking between BSA and myoglobin which was confirmed by SDS-PAGE and subsequent amino acid sequencing. Moreover, dityrosine was formed in the initial phases of the reaction for all concentrations of H2O2. However, initially formed dityrosine was subsequently utilized in reactions employing stoichiometric excess of H2O2 to MMb. The observed breakdown of dityrosine was ascribed to additional radical species formed from the interaction between H2O2 and the hyper-valent iron-center of H2O2-activated MMb.
authors.affiliation: Department of Animal Product Quality, Danish Institute of Agricultural Sciences, Research Center Foulum, Tjele. henrik.ostdal@agrsci.dk
authors.forename: H
authors.initials: H
authors.lastname: Ostdal
authors.forename: S G
authors.initials: SG
authors.lastname: Søgaard
authors.forename: E
authors.initials: E
authors.lastname: Bendixen
authors.forename: H J
authors.initials: HJ
authors.lastname: Andersen
mesh_terms: D000818:Animals; D002417:Cattle; D004578:Electron Spin Resonance Spectroscopy; D006736:Horses; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007700:Kinetics; D008786:Metmyoglobin / Q000378:metabolism*; D009211:Myoglobin / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism*; D013997:Time Factors; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: D009211:Myoglobin; D008786:Metmyoglobin; D012710:Serum Albumin, Bovine; D014443:Tyrosine; D006861:Hydrogen Peroxide; C007543:dityrosine