Proteomics of oxidative stress in mice

Aphrodite Vasilaki, Deborah Simpson, Francis McArdle, Lynne McLean, Robert J. Beynon, Holly Van Remmen, Arlan G. Richardson, Anne McArdle, John A Faulkner and Malcolm J. Jackson (2007) Formation of 3-nitrotyrosines in carbonic anhydrase III is a sensitive marker of oxidative stress in skeletal muscle PROTEOMICS – Clinical Applications 1: 362–372 [PDF]

Oxidation of skeletal muscle proteins has been reported to occur following contractions, with ageing, and with a variety of disease states, but the nature of the oxidised proteins has not been identified. A proteomics approach was utilised to identify major proteins that contain carbonyls and/or 3-nitrotyrosine (3-NT) groups in the gastrocnemius (GTN) muscles of adult (5– 11 months of age) and old (26–28 months of age) wild type (WT) mice and adult mice lacking copper, zinc superoxide dismutase (Sod12/2 mice), manganese superoxide dismutase (Sod21/2 mice) or glutathione peroxidase 1 (GPx12/2 mice). In quiescent GTN muscles of adult and old WT mice, protein carbonylation and/or formation of 3-NT occurred in several proteins involved in glycolysis, as well as creatine kinase and carbonic anhydrase III. Following contractions, the 3-NT intensity was increased in specific protein bands from GTN muscles of both adult and old WT mice. In quiescent GTN muscles from adult Sod12/2, Sod21/2 or GPx12/2 mice compared with age-matched WT mice only carbonic anhydrase III showed a greater 3-NT content. We con- clude that formation of 3-NT occurs readily in response to oxidative stress in carbonic anhydrase III and this may provide a sensitive measure of oxidative damage to muscle proteins.