2‐D DIGE analysis of the mitochondrial proteome from human skeletal muscle reveals time course‐dependent remodelling in response to 14 consecutive days of endurance exercise training

Abstract

Adaptation of skeletal muscle to repeated bouts of endurance exercise increases aerobic capacity and improves mitochondrial function. However, the adaptation of human skeletal muscle mitochondrial proteome to short-term endurance exercise training has not been investigated. Eight sedentary males cycled for 60 min at 80% of peak oxygen consumption (VO 2peak ) each day for 14 consecutive days, resulting in an increase in VO 2peak of 17.573.8% (po0.01). Mitochondria-enriched protein fractions from skeletal muscle biopsies taken from m. vastus lateralis at baseline, and on the morning following the 7th and 14th training sessions were subjected to 2-D DIGE analysis with subsequent MS followed by database interrogation to identify the proteins of interest. Thirty-one protein spots were differentially expressed after either 7 or 14 days of training (ANOVA, po0.05). These proteins included subunits of the electron transport chain, enzymes of the tricarboxylic acid cycle, phosphotransfer enzymes, and regulatory factors in mitochondrial protein synthesis, oxygen transport, and antioxidant capacity. Several proteins demonstrated a time course-dependent induction during training. Our results illustrate the phenomenon of skeletal muscle plasticity with the extensive remodelling of the mitochondrial proteome occurring after just 7 days of exercise training suggestive of enhanced capacity for adenosine triphosphate generation at a cellular level.

The proteomic signature of insulin-resistant human skeletal muscle reveals increased glycolytic and decreased mitochondrial enzymes

“…There have been several previous approaches to the study of the human skeletal muscle proteome [13,14] and its alterations in insulin-resistant muscle [15][16][17][18]. Two studies have used the two-dimensional gel approach [15,16].…”

“…Within the last years, the combination of twodimensional differential gel electrophoresis (2D-DIGE) and fluorescence staining of proteins followed by protein identification using high-sensitivity MS/MS methods has proven to be a suitable gel-based method for the quantification of changes in abundance of proteins or patterns of proteins in human tissues and has recently been used to compare the human skeletal muscle mitochondrial proteome before and after endurance exercise training [14]. Here, we applied this technology to investigate alterations of the skeletal muscle proteome in obesity and type-2-diabetesassociated insulin resistance, respectively.…”

Exercise Proteomics: Review

“…The physiological conditioning of fatigue resistance and the bio-energetic enhancement of aerobic performance are based on finely tuned physiological machinery that promotes endurance performance (Seene et al2011). Proteomic profiling of the effect of endurance training on human vastus lateralis muscle revealed distinct adaptations in the expression profile of the mitochondrial proteome, especially affecting enzymes such as NADH dehydrogenase and ATP synthase (Egan et al 2011). The mitochondria-enriched fraction from skeletal muscle biopsies showed differential expression patterns of enzymes of the citric acid cycle, oxidative phosphorylation, mitochondrial protein synthesis, oxygen transportation and antioxidant capacity following endurance training (Egan et al 2011).…”

“…Proteomic profiling of the effect of endurance training on human vastus lateralis muscle revealed distinct adaptations in the expression profile of the mitochondrial proteome, especially affecting enzymes such as NADH dehydrogenase and ATP synthase (Egan et al 2011). The mitochondria-enriched fraction from skeletal muscle biopsies showed differential expression patterns of enzymes of the citric acid cycle, oxidative phosphorylation, mitochondrial protein synthesis, oxygen transportation and antioxidant capacity following endurance training (Egan et al 2011). A variety of proteomic surveys with animal models of endurance training have confirmed exercise-induced mitochondrial remodelling and an increased capacity for oxidative metabolism.…”

“…Exercise proteomics was used to study protein alterations in humans (Malm et al2012;Egan et al 2011;Norheim et al 2011;Hody et al 2011;Moriggi et al 2010;Holloway et al 2009) and animal models of physical activity Magherini et al 2012;Macedo et al 2012;Yamaguchi et al 2010;Bouwman et al 2010;Burniston 2008;Donoghue et al 2007;Guelfi et al 2006;Donoghue et al 2005). The neuromuscular system has a vast capacity to become accustomed to a great variety of physical demands and differing training conditions by muscle remodelling involving changes in contractile properties, metabolic pathways and tissue mass (Bassel and Olson 2006).…”