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REDOX PROTEOMICS OF MOUSE SKELETAL MUSCLE AGEING
S42
E. Chiaradia et al. / Free Radical Biology and Medicine 96 (2016) S32–S69
modifications of lipid, protein and DNA were significantly enhanced. It indicates that URB597 enhances disturbances in crosstalk between endocannabinoids, oxidants and inflammatory factors what may lead to functional disorders in the kidney of hypertensive rats. http://dx.doi.org/10.1016/j.freeradbiomed.2016.04.085 P-23
Redox proteomics of mouse skeletal muscle ageing Neil T. Smith, Malcolm J. Jackson, Brian McDonagh MRC-Arthritis Research UK Centre for Integrated Research into Musculoskeletal Aging (CIMA), Department of Musculoskeletal Biology, Institute of Aging and Chronic Disease, University of Liverpool, Liverpool, UK
Reactive oxygen species (ROS) are increasingly recognised as crucial signalling molecules required for normal cellular functioning. Reversible redox modifications of regulatory amino acids induced by ROS offers a dynamic and versatile means to rapidly alter the activity or structure of proteins in response to external and internal stimuli. Skeletal muscle generates ROS during contractions and muscle ageing is accompanied by an altered redox environment within muscle fibres, but it is unknown whether these changes are universal across all muscle types. Aims: To investigate changes in the redox environment with age we used two metabolically different muscle tissues, quadriceps and soleus from male C57BL/6 adult (12 months) and old (24 months) mice. We utilised a redox proteomic approach combining label free proteomics and differential Cys labelling of reduced and reversibly oxidised Cys residues to identify how these muscles change with age. This approach allowed a global overview of the proteomic changes with age but also the relative quantification of the oxidation state of redox sensitive Cys residues in the context of the protein's abundance. Results: Label free analysis of soleus muscle revealed a significant down regulation of mitochondrial proteins with age, while the quadriceps showed a significant increase in cytoskeletal proteins and a significant decrease in glycolytic enzymes with age. Several of the metabolically active proteins were further investigated with western blotting and enzyme activity analysis. Differential labelling of specific Cys residues in both quadriceps and soleus tissues revealed a decrease in the number of redox sensitive proteins detected with age. These results indicate that skeletal muscles respond differently to the effects of ageing, reflecting the underlying metabolic processes within the tissue. http://dx.doi.org/10.1016/j.freeradbiomed.2016.04.086
Gerasimos Terzis 4, Apostolos Sovatzidis 5, Maria Protopappa 3, Konstantinos Papanikolaou 1, Alexandra Avloniti 3 1
School of Physical Education and Sports Sciences, University of Thessaly, Trikala, Greece 2 National Hellenic Research Foundation, Athens, Greece 3 School of Physical Education and Sports Sciences, Democritus University of Thrace, Komotini, Greece 4 School of Physical Education and Sports Sciences, University of Ahens, Athens, Greece 5 Medical School, Unit of Nephrology, Democritus University of Thrace, Alexandroupolis, Greece Protein supplementation has been well-described to give rise to skeletal muscle protein synthesis. When resistance exercise precedes protein injestion the anabolic effect is synergestic leading to a positive, net protein balance. Following, eccentric exercise, it is evident the aseptic skeletal muscle microtrauma, accompanied by increased oxidative stress and inflammatory responses. Protein supplementation has been shown to reduce oxidative stress and inflammation, in clinical cnoditions. Aims: The objective was to examine the effect of protein supplementation on redox status and redox-sensitive signaling pathways, following eccentric exercise-induced skeletal muscle microtrauma. Methods: In a double-blind, crossover design, 12 young males, received placebo (Pla) or milk protein isolate (PRO:80% casein & 20% whey protein), following unilateral, eccentric exercise (300 ecentric contraction) on isokinetic dynamometer. Participnats in PRO received 20 g of protein every 3 h in the exercise day (on four occasions, 80 g in total) and a bolus of 20 g of protein daily with breakfast on the remaining 7 days. In each condition, muscle performance was evaluated before exercise, 2 h post-exercise, and daily for 7 consecutive days. Muscle biopsy samples from vastus lateralis and blood samples were collected before exercise as well as on day 2 and day 8.
Results: PRO attenuated the phosphorylation of nuclear factor kB and strength decline during recovery. PRO also enhanced the elevation of mammalian target of rapamycin (mTOR) and ribosomal protein S6. Moreover, PRO preserved GSH levels maintaining a higher GSH/GSSG ratio, compared to Pla. Proteasome activity was also preserved by protein supplementation whereas no changes observed at the proteasome's protein expression level. http://dx.doi.org/10.1016/j.freeradbiomed.2016.04.087 P-25
Redox status in response to exercise training in heavy drinkers
P-24
Protein supplementation alters redox-status and proteosomic activity following aseptic inflammation induced by exercise Ioannis G. Fatouros 1, Dimitrios Draganidis 1, Niki Chondrogianni 2, Athanasios Z. Jamurtas 1, Athanasios Chatzinikolaou 3,
Athanasios Z. Jamurtas 1,2, Kalliopi Georgakouli 1,2,3, Eirini Manthou 1,2, Ioannis Fatouros 1,2, Yannis Theodorakis 1 1
Department of Physical Education & Sport Science, University of Thessaly, Trikala, Greece 2 Department of Kinesiology, Institute for Research and Technology Thessaly, Trikala, Greece
E. Chiaradia et al. / Free Radical Biology and Medicine 96 (2016) S32–S69
modifications of lipid, protein and DNA were significantly enhanced. It indicates that URB597 enhances disturbances in crosstalk between endocannabinoids, oxidants and inflammatory factors what may lead to functional disorders in the kidney of hypertensive rats. http://dx.doi.org/10.1016/j.freeradbiomed.2016.04.085 P-23
Redox proteomics of mouse skeletal muscle ageing Neil T. Smith, Malcolm J. Jackson, Brian McDonagh MRC-Arthritis Research UK Centre for Integrated Research into Musculoskeletal Aging (CIMA), Department of Musculoskeletal Biology, Institute of Aging and Chronic Disease, University of Liverpool, Liverpool, UK
Reactive oxygen species (ROS) are increasingly recognised as crucial signalling molecules required for normal cellular functioning. Reversible redox modifications of regulatory amino acids induced by ROS offers a dynamic and versatile means to rapidly alter the activity or structure of proteins in response to external and internal stimuli. Skeletal muscle generates ROS during contractions and muscle ageing is accompanied by an altered redox environment within muscle fibres, but it is unknown whether these changes are universal across all muscle types. Aims: To investigate changes in the redox environment with age we used two metabolically different muscle tissues, quadriceps and soleus from male C57BL/6 adult (12 months) and old (24 months) mice. We utilised a redox proteomic approach combining label free proteomics and differential Cys labelling of reduced and reversibly oxidised Cys residues to identify how these muscles change with age. This approach allowed a global overview of the proteomic changes with age but also the relative quantification of the oxidation state of redox sensitive Cys residues in the context of the protein's abundance. Results: Label free analysis of soleus muscle revealed a significant down regulation of mitochondrial proteins with age, while the quadriceps showed a significant increase in cytoskeletal proteins and a significant decrease in glycolytic enzymes with age. Several of the metabolically active proteins were further investigated with western blotting and enzyme activity analysis. Differential labelling of specific Cys residues in both quadriceps and soleus tissues revealed a decrease in the number of redox sensitive proteins detected with age. These results indicate that skeletal muscles respond differently to the effects of ageing, reflecting the underlying metabolic processes within the tissue. http://dx.doi.org/10.1016/j.freeradbiomed.2016.04.086
Gerasimos Terzis 4, Apostolos Sovatzidis 5, Maria Protopappa 3, Konstantinos Papanikolaou 1, Alexandra Avloniti 3 1
School of Physical Education and Sports Sciences, University of Thessaly, Trikala, Greece 2 National Hellenic Research Foundation, Athens, Greece 3 School of Physical Education and Sports Sciences, Democritus University of Thrace, Komotini, Greece 4 School of Physical Education and Sports Sciences, University of Ahens, Athens, Greece 5 Medical School, Unit of Nephrology, Democritus University of Thrace, Alexandroupolis, Greece Protein supplementation has been well-described to give rise to skeletal muscle protein synthesis. When resistance exercise precedes protein injestion the anabolic effect is synergestic leading to a positive, net protein balance. Following, eccentric exercise, it is evident the aseptic skeletal muscle microtrauma, accompanied by increased oxidative stress and inflammatory responses. Protein supplementation has been shown to reduce oxidative stress and inflammation, in clinical cnoditions. Aims: The objective was to examine the effect of protein supplementation on redox status and redox-sensitive signaling pathways, following eccentric exercise-induced skeletal muscle microtrauma. Methods: In a double-blind, crossover design, 12 young males, received placebo (Pla) or milk protein isolate (PRO:80% casein & 20% whey protein), following unilateral, eccentric exercise (300 ecentric contraction) on isokinetic dynamometer. Participnats in PRO received 20 g of protein every 3 h in the exercise day (on four occasions, 80 g in total) and a bolus of 20 g of protein daily with breakfast on the remaining 7 days. In each condition, muscle performance was evaluated before exercise, 2 h post-exercise, and daily for 7 consecutive days. Muscle biopsy samples from vastus lateralis and blood samples were collected before exercise as well as on day 2 and day 8.
Results: PRO attenuated the phosphorylation of nuclear factor kB and strength decline during recovery. PRO also enhanced the elevation of mammalian target of rapamycin (mTOR) and ribosomal protein S6. Moreover, PRO preserved GSH levels maintaining a higher GSH/GSSG ratio, compared to Pla. Proteasome activity was also preserved by protein supplementation whereas no changes observed at the proteasome's protein expression level. http://dx.doi.org/10.1016/j.freeradbiomed.2016.04.087 P-25
Redox status in response to exercise training in heavy drinkers
P-24
Protein supplementation alters redox-status and proteosomic activity following aseptic inflammation induced by exercise Ioannis G. Fatouros 1, Dimitrios Draganidis 1, Niki Chondrogianni 2, Athanasios Z. Jamurtas 1, Athanasios Chatzinikolaou 3,
Athanasios Z. Jamurtas 1,2, Kalliopi Georgakouli 1,2,3, Eirini Manthou 1,2, Ioannis Fatouros 1,2, Yannis Theodorakis 1 1
Department of Physical Education & Sport Science, University of Thessaly, Trikala, Greece 2 Department of Kinesiology, Institute for Research and Technology Thessaly, Trikala, Greece