Profiling Carbonylated Proteins in Heart and Skeletal Muscle Mitochondria from Trained and Untrained Mice

Abstract

Understanding the relationship between physical exercise, reactive oxygen species, and skeletal muscle modification is important in order to better identify the benefits or the damages that appropriate or inappropriate exercise can induce. Heart and skeletal muscles have a high density of mitochondria with robust energetic demands, and mitochondria plasticity has an important role in both the cardiovascular system and skeletal muscle responses. The aim of this study was to investigate the influence of regular physical activity on the oxidation profiles of mitochondrial proteins from heart and tibialis anterior muscles. To this end, we used the mouse as animal model. Mice were divided into two groups: untrained and regularly trained. The carbonylated protein pattern was studied by two-dimensional gel electrophoresis followed by Western blot with anti-dinitrophenyl hydrazone antibodies. Mass spectrometry analysis allowed the identification of several different protein oxidation sites, including methionine, cysteine, proline, and leucine residues. A large number of oxidized proteins were found in both untrained and trained animals. Moreover, mitochondria from skeletal muscles and heart showed almost the same carbonylation pattern. Interestingly, exercise training seems to increase the carbonylation level mainly of mitochondrial proteins from skeletal muscle.

Effect of Functional Fitness on Plasma Oxidation Level in Elders: Reduction of the Plasma Oxidants and Improvement of the Antioxidant Barrier

“…Both plasma and saliva extracts from T0 and T1 groups were separately suspended in 8 M urea, 4% 3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), 65 mM dithiothreitol (DTT). Proteins were quantified through Bradford colorimetric method (Biorad, Hercules, California, USA) and 80 µg were separated on 11 cm immobilized pH gradient (IPG) strips (pH 3-10 NL) on a PROTEAN i12 IEF System (BIO-RAD) as described in Carpentieri et al, 2016 [18]. Next, the strips were incubated in 2 N HCl with 10 mM 2,4-dinitrophenyl-hydrazine, DNPH (Sigma, St. Louis, MO) for 20 min at 25°C, to enable the derivatization of protein-bound carbonyls as reported in Carpentieri et al 2016 [18].…”

Irreversible oxidative post-translational modifications in heart disease

“…Mitochondria mouse heart and TCA cycle, amino acid metabolism, respiratory chain Carpentieri, A, et al [75].…”

Data on protein abundance alteration induced by chronic exercise in mdx mice model of Duchenne muscular dystrophy and potential modulation by apocynin and taurine

“…The in vivo weakness produced by such a protocol is observed exclusively in mdx mice with no similar effects in wild type mice [4] , [5] . In fact, protocols used to induce training effects in wild types mice usually consist of continuous running at 20 m/min for at least 15 min using a treadmill slope of 10°, five days a week, for eight weeks [6] . To exclude training effects in wt animals we checked the amount of selected proteins in wt animals subjected to the same exercise protocol of mdx mice.…”