title: Effects of manipulation of the caspase system on myofibrillar protein degradation in vitro.
abstract: Apoptosis via the intrinsic caspase 9 pathway can be induced by oxidative stressors hydrogen peroxide (H₂O₂) and N-(4 hydroxyphenol) rentinamide (fenretinide), a synthetic retinoid. Accelerated muscle atrophy and proteolysis in muscle-wasting conditions have been linked to oxidative stress and activated protease systems. Therefore, the hypothesis of this study was that proteolysis of myofibrillar proteins could be manipulated through the induction or inhibition of the caspase system. After slaughter, LM and supraspinatus muscles from callipyge (n = 5) and normal (n = 3) lambs were excised, finely diced, and incubated with treatment buffers containing oxidative stressors fenretinide or H₂O₂, recombinant caspase 3, caspase-specific inhibitor N-acetyl-Asp-Glu-Val-Asp-CHO (DEVD), or control solution. Muscle samples were incubated for 1, 2, 7, and 21 d at 4°C. Activation of the initiator caspase, caspase 9, and myofibrillar protein degradation was determined by SDS-PAGE and Western blotting. Results showed that fenretinide, H₂O₂, and recombinant caspase 3 increased (P < 0.05) proteolysis of myofibril proteins, whereas DEVD inhibited degradation (P < 0.05). Proteolysis of myofibrillar proteins increased with incubation time (P < 0.0001), and incubation time × treatment interactions (P < 0.05) indicated that the treatment effects did not all occur at the same rate. This study has shown that manipulation of the caspase system through induction or inhibition of activity can affect degradation of myofibrillar proteins, providing further evidence that the caspase system could be involved in postmortem proteolysis and tenderization. However, these stimulated changes were not sufficient to overcome the lack of proteolysis that is characteristic of muscle from callipyge lambs.
authors.affiliation: Roman L. Hruska US Meat Animal Research Center, USDA, ARS, Clay Center, NE 68933-0166, USA.
authors.forename: C M
authors.initials: CM
authors.lastname: Kemp
authors.forename: T L
authors.initials: TL
authors.lastname: Wheeler
mesh_terms: D000818:Animals; D020169:Caspases / Q000378:metabolism*; D005260:Female; D005838:Genotype; D018482:Muscle, Skeletal / Q000201:enzymology*; D009210:Myofibrils / Q000378:metabolism*; D011489:Protein Denaturation; D012756:Sheep / Q000235:genetics / Q000502:physiology*
chemical_list: D020169:Caspases