calpastatin and 3-methylhistidine

This experiment was conducted to determine the relationship between 3-methylhistidine (3MH) production and proteinase activity in skeletal muscles of growing barrows. Barrows at 13 wk of age were randomly assigned to either control diet available on an ad libitum basis (21% of ME consisted of protein; control group), control diet fed restricted (pair-fed with barrows in protein-free group; intake-restricted group), or protein-free diet available on an ad libitum basis (protein-free group) for 14 d. During the last 3 d, blood samples were collected for determination of 3MH production rate, which is a measure of myofibrillar protein breakdown. At slaughter, two muscles were taken: masseter (M) and longissimus (L) muscles. The muscle samples were analyzed for calpastatin, mu-calpain, m-calpain, multicatalytic proteinase (MCP), cathepsin B, cathepsins B+L, and cystatins activities. Both muscles were also analyzed for amounts of DNA, RNA, total protein, and myofibrillar and sarcoplasmic proteins. Growth rate (kilograms/day) was influenced by dietary treatments (P < .05). Fractional breakdown rate (FBR, percentage/day) of skeletal muscle, as calculated from 3MH production rate (micromoles.kilogram-1.day-1), was 27% higher for the protein-free group than for the control group. However, no differences in proteinase activities were observed, except for lower MCP activity in the M muscle of the protein-free group than in that of the other groups (P < .05). In the present study, no direct relation was observed between myofibrillar protein degradation rate and proteinase activities in skeletal muscle during a protein-free feeding strategy.

Topics: Animals; Calcium-Binding Proteins; Calpain; Cathepsins; Cystatins; Diet, Protein-Restricted; Dietary Proteins; DNA; Endopeptidases; Liver; Male; Methylhistidines; Muscle Proteins; Muscle, Skeletal; Organ Size; Protein Deficiency; Random Allocation; RNA; Swine; Swine Diseases

The objectives were to investigate the mechanisms by which glucocorticoids control proteolysis in muscle cells and the relationship between the calpain:calpastatin system and proteolysis in muscle. Female rabbits were treated with 1 mg dexamethasone (Dex)/kg body weight per day for 0, 1, 2 or 4 days after which animals were killed and muscle samples taken for analyses. Dex reduced urinary N tau-methylhistidine (NMH) 48% (day 4 versus day 1 of Dex treatment) and muscle NMH concentrations by 49% (day 1) to 40% (day 2) respectively, suggesting that protein degradation was reduced. To investigate whether the changes in apparent proteolysis were related to calpains, we examined the effects of Dex on muscle calpain and calpastatin activities. These were unaffected by Dex. This implies that Dex-dependent changes in degradation are not mediated by changes in muscle calpain or calpastatin activities. We studied the effects of Dex on calpain and calpastatin gene expression as a means of clarifying the relationships between proteinase gene expression and proteinase activities. mu-Calpain mRNA concentration was unaffected by Dex but m-calpain mRNA and calpastatin mRNA concentrations were reduced by 42-55% and 40% respectively. Dex had a similar effect on beta-actin mRNA. Although calpain and calpastatin genes behaved as house-keeping genes, changes in their expression mimicked apparent changes in proteolysis. The observation that calpain and calpastatin activities were unchanged indicates that additional regulation of the calpain:calpastatin system exists at other sites in muscle cells. To determine whether Dex-dependent changes in proteolysis were mediated indirectly, we assayed the effects of Dex on plasma thyroid hormone concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Blotting, Northern; Calcium-Binding Proteins; Calpain; Dexamethasone; Female; Gene Expression; Homeostasis; Methylhistidines; Muscles; Rabbits; RNA; Thyroxine; Triiodothyronine

The effect of castration on endogenous proteinase activity and myofibrillar protein turnover was investigated in cattle. Six each of MARC III composite bulls and steers weighing approximately 210 kg were given ad libitum access to a typical growing diet. At 0, 42, 84, 126, and 168 d, two consecutive 24-h urine samples were obtained. Urine was analyzed for N tau-methylhistidine (N tau MH) and creatinine. Following slaughter after 170 d on feed, a longissimus muscle sample was removed immediately from each carcass for quantification of mu-calpain, m-calpain, calpastatin, cystatin(s), cathepsin B, and cathepsin B + L activities. Bulls were heavier (P < .05) at 126 and 168 d and more efficient (P < .05) in conversion of feed to gain at 84 and 168 d than were steers. Compared with steers, bulls excreted less (P < .05) N tau MH at 84, 126, and 168 d and displayed lower (P < .05) fractional degradation rates (FDR) at all sample times. No differences (P > .05) in calpain or cathepsin activities were observed between bulls and steers. However, muscle from bulls had greater (P < .05) activities of calpastatin and cystatin(s) than that from steers. A negative relationship existed between d-168 FDR and calpastatin (r = -.72; P < .05) and cystatin (r = -.62; P < .05) activities. These results indicate that decreased FDR of skeletal muscle from growing bulls contributes to their greater efficiency of growth and could be related partially to cystatin-mediated cathepsin activity and(or) calpastatin-mediated calpain activity.

Topics: Animals; Calcium-Binding Proteins; Calpain; Cathepsins; Cattle; Creatinine; Cystatins; DNA; Endopeptidases; Male; Methylhistidines; Muscle Development; Muscle Proteins; Muscles; Orchiectomy; Random Allocation; RNA; Weight Gain

The objectives of this study were to examine effects of a beta-adrenergic agonist (cimaterol) on growth and muscle development in rabbits and to examine cimaterol's effects on myofibrillar protein degradation (MPD) and on activities of several proteolytic enzymes including the calcium-dependent proteinases (CDP). Twelve New Zealand White rabbits were assigned to either control diets or to diets containing cimaterol for 35 d, after which they were killed and effects on performance and tissue weight gains were determined. Urine was collected from d 21 through 28 from each rabbit for assessment of N tau-methylhistidine (NMH) excretion. Cimaterol increased rates of gain, efficiency of gain and skeletal muscle weights. Enhancement in muscle weight was associated with an increase in total DNA and with a reduction in NMH. Cimaterol did not affect activities of cathepsin B, cathepsin D or neutral serine proteinase, but it reduced activities of the millimolar and micromolar forms of the CDP by 58 and 57%, respectively, and it reduced activity of the inhibitor of the CDP (calpastatin) by 52%. Cimaterol-dependent myofibrillar protein accretion was likely mediated, at least in part, by a reduction in MPD. The change in MPD was associated with a reduction in muscle CDP activities. Cimaterol-dependent muscle hypertrophy therefore may involve changes in calcium-dependent proteolysis of myofibrillar proteins. The significance of the effects of cimaterol on calpastatin activity is not known.

Topics: Animals; Calcium-Binding Proteins; Calpain; DNA; Ethanolamines; Liver; Male; Methylhistidines; Muscle Development; Muscle Proteins; Muscles; Myofibrils; Rabbits; Random Allocation; RNA; Weight Gain