Radical-induced damage to bovine serum albumin: role of the cysteine residue

• Published in: Free radical research communications Vol. 18; no. 6; p. 353
• Main Authors: Davies, M J, Gilbert, B C, Haywood, R M
• Format: Journal Article
• Language: English
• Published: Switzerland 1993
• Subjects: Cyclic N-Oxides; Cysteine; Electron Spin Resonance Spectroscopy - methods; Free Radicals; Hydroxides; Hydroxyl Radical; Kinetics; Serum Albumin, Bovine - chemistry; Spin Labels
• ISSN: 8755-0199
• DOI: 10.3109/10715769309147502

The reactions of cerium(IV) and the hydroxyl radical [generated from iron(ii)/H2O2] with bovine serum albumin (BSA) have been investigated by EPR spin trapping. With the former reagent a protein-derived thiyl radical is selectively generated; this has been characterized via the anisotropic EPR spectra observed on reaction of this radical with the spin trap DMPO. Blocking of the thiol group results in the loss of this species and the detection of a peroxyl radical, believed to be formed by reaction of oxygen with initially-generated, but undetected, carbon-centred radicals from aromatic amino acids. Experiments with a second spin trap (DBNBS) confirm the formation of these carbon-centred species and suggest that damage can be transferred from the thiol group to carbon sites in the protein. A similar transfer pathway can be observed when hydroxyl radicals react with BSA. Further experiments demonstrate that the reverse process can also occur: when hydroxyl radicals react with BSA, the thiol group appears to act as a radical sink and protects the protein from denaturation and fragmentation through the transfer of damage from a carbon site to the thiol group. Thiol-blocked BSA is shown to be more susceptible to damage than the native protein in both direct EPR experiments and enzyme digestion studies. Oxygen has a similar effect, with more rapid fragmentation detected in its presence than its absence.