Oxidation of porcine Myosin by hypervalent myoglobin: the role of thiol groups.
Oxidation of the myofibrillar muscle protein myosin from pork by hypervalent myoglobin species (MbFe(III)/H 2O2 radical generating system) was investigated in aqueous solution in the pH range of 5.0-7.8 by electron spin resonance (ESR) spectroscopy using N- tert-butyl-alpha-phenylnitrone (PBN) as spin trap and indirectly by determination of the rate of reduction of hypervalent myoglobin species by UV spectroscopy. Cross-linking of myosin was examined by SDS-PAGE. The target for oxidative modification of myosin was studied by thiol blocking by N-acetylmaleimide (NEM) and by determining oxidative modification of myosin thiols. The reaction between myosin and hypervalent myoglobin was fast and showed little dependence on pH. The myosin radicals formed were observed to be short-lived. Myosin thiols are suggested to be the main target for oxidative modification, as NEM-treated myosin did not form radicals in the presence of hypervalent myoglobin. A significant decrease in thiol content was already demonstrated 25 s after initiation of oxidation of myosin. The majority of myosin heavy chain (MHC) was demonstrated to be cross-linked through intermolecular disulfide bonding 1 h after initiation of oxidation. This demonstrates that thiols are important for radical formation and cross-linking of myosin during oxidation with hypervalent myoglobin at the pH of meat products.
Frederiksen
Anne M
AM
Food Chemistry, Department of Food Science, Faculty of Life Sciences, University of Copenhagen, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark.
Lund
Marianne N
MN
Andersen
Mogens L
ML
Skibsted
Leif H
LH
D000818:Animals; D003432:Cross-Linking Reagents; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008460:Meat / Q000032:analysis*; D009211:Myoglobin / Q000737:chemistry*; D009218:Myosins / Q000737:chemistry*; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D013438:Sulfhydryl Compounds / Q000032:analysis*; D013552:Swine
D003432:Cross-Linking Reagents; D005609:Free Radicals; D009211:Myoglobin; D013438:Sulfhydryl Compounds; D009218:Myosins