Fractional Rates Of Protein Degradation Research Articles

337 Efficient RFI Bulls Presented Lower Fractional Synthesis Rates of Plasma Proteins When Fed Corn but Not Grass Based Diets

Abstract Protein turnover (PT), the continual synthesis and degradation of body proteins not leading to protein gain, is an essential high energy-demanding process. We assumed that PT might explain the between-animal variations of residual feed intake (RFI). The objective was to measure PT in extreme RFI cattle fed two contrasted diets (grass or corn-based). We conducted a RFI test for 84 days with 100 Charolais bulls and we selected the 32 most extreme (8 per diet and RFI group) for PT measurements using 1) the urinary 3-methyl-histidine to creatinine ratio, as a biomarker of the fractional protein degradation rate (FDR) of skeletal-muscle and 2) the isotopic N turnover rate measured in urine and plasma, as a proxy, respectively, of the whole-body FDR and the fractional protein synthesis rates (FSR) of plasma proteins. The 3-methyl-histidine and creatinine were determined from 10 d total urine collection. Isotopic N turnover in urine and plasma was evaluated by modelling the 15N depletion rate over 112 d following an isotopic N dietary change. Higher plasma FSR and higher skeletal-muscle and whole-body FDR were observed with corn-vs-grass diets (≥11%; P ≤ 0.03), in line with higher metabolizable protein and net energy intakes (≥10%, P = 0.001). Differences between extreme RFI animals were noted with the corn diets only, where efficient animals presented significant lower plasma FSR (-10%; P = 0.04) and numerically lower skeletal-muscle and whole-body FDR (-13% and - 8.9%; P > 0.16 respectively) than non-efficient. Non-significant differences were probably due to an insufficient size of our experimental setup. Plasma FSR is related to the PS of hepatic exportation, hence the lower plasma FSR observed in efficient RFI animals fed corn diets may reflect a lower organs to carcass ratio. Altogether results suggests that efficient RFI bulls fed corn diets had a lower hepatic PT with no-significant changes of whole-body and skeletal muscle PT.

The IGF-1/Akt/S6 Signaling Pathway is Age-Dependently Downregulated in the Chicken Breast Muscle.

The skeletal muscle mass is known to be controlled by the balance between protein synthesis and degradation. The fractional rate of protein synthesis has been reported to decrease age-dependently from 1 to 4 weeks of age in the chicken breast muscle (pectoralis major muscle). On the other hand, age-dependent change of the fractional protein degradation rate was reported to be less in the skeletal muscle of chickens. These findings suggest that protein synthesis is age-dependently downregulated in chicken muscle. We herein investigated the age-dependent changes in protein synthesis or proteolysis-related factors in the breast muscle of 7, 14, 28, and 49-day old broiler chickens. IGF-1 mRNA level, phosphorylation rate of Akt, and phospho-S6 content were coordinately decreased in an age-dependent manner, suggesting that IGF-1-stimulated protein synthesis is downregulated with age in chicken breast muscle. In contrast, atrogin-1, one of the proteolysis-related factors, gradually increased with age at mRNA levels. However, plasma Nτ-methylhistidine concentration, an indicator of skeletal muscle proteolysis, did not coordinately change with atrogin-1 mRNA levels. Taken together, our results suggest that the IGF-1/Akt/S6 signaling pathway is age-dependently downregulated in the chicken breast muscle.

In vivo measurements of the contributions of protein synthesis and protein degradation in regulating cardiac pressure overload hypertrophy in the mouse

Cardiac hypertrophy is generated in response to hemodynamic overload by altering steady-state protein metabolism such that the rate of protein synthesis exceeds the rate of protein degradation. To determine the relative contributions of protein synthesis and degradation in regulating cardiac hypertrophy in mice, a continuous infusion strategy was developed to measure myocardial protein synthesis rates in vivo. Osmotic mini-pumps were implanted in the abdominal cavity to infuse radiolabeled leucine in mice that are conscious and ambulatory. Protein synthesis rates were calculated by measuring incorporation of leucine into myocardial protein over 24 h prior to each time point and dividing by the specific radioactivity of plasma leucine. Compared to sham-operated controls, fractional rates of protein synthesis (K(s)) increased significantly at days 1 and 3 of TAC, but was lower on day 7 and returned to control values by day 14. These changes coincided with the curvilinear increase in LV mass that characterizes the hypertrophic response. Fractional rates of protein degradation (K(d)) were calculated by subtracting the rate of myocardial growth from the corresponding K(s) value. K(d) fell at days 1 and 3 of TAC, increased at day 7 and returned to control on day 14. Thus, the increase in LV mass generated in response to pressure overload is caused by acceleration of K(s) and suppression of K(d). As the growth rate slows, a new steady-state is achieved once the hypertrophic response is completed.

Maslinic acid added to the diet increases growth and protein-turnover rates in the white muscle of rainbow trout ( Oncorhynchus mykiss)

Maslinic acid (2-α, 3-β-dihydroxiolean-12-en-28-oic acid) is a triterpenoid compound present in fruit and leaves of Olea europaea that can be used as an additive in the diet of trout. The present work investigates the effects of maslinic acid on growth, protein-turnover rates and nucleic acid concentration in trout white muscle. Five groups of 180 trout of a mean body mass of 20 g were fed for 225 days with diets containing 0, 1, 5, 25 and 250 mg of maslinic acid per kg of diet. At the end of the experiment, white-muscle weight and protein-accumulation rate of trout fed with maslinic acid were higher than in control. The total content of DNA, RNA, and protein in trout fed with 25 and 250 mg of maslinic acid kg − 1 were significantly higher than in control. The protein:DNA ratio was also slightly higher than control. In the same groups of trout, fractional ( K S) and absolute ( A S) protein-synthesis rates increased to more than 80% over the control values while no differences were found in the fractional protein-degradation rate ( K D). These results, similar to previous findings in liver, show that maslinic acid can act as a growth factor when added to a standard trout diet.

Growth, carcass quality, and protein and energy metabolism in beef cattle with different growth potentials and residual feed intakes1,2

Twenty-four beef steers (predominantly Angus x Hereford, 14 to 18 mo of age, 403 +/- 3 kg of BW), were housed and fed in individual pens for about 122 d. Twelve steers came from a herd that had been selected for growth (high growth; HG) and the other 12 from a herd with no selection program (low growth; LG). Another 6 steers (3 from each group) were slaughtered at the beginning to obtain the initial composition. All steers were fed the same corn-based diet (3.06 Mcal of ME/kg of DM, 13.6% CP) on an ad libitum basis. Two weeks before slaughter, total urine was collected for 5 d for estimation of 3-methylhistidine excretion and myofibrillar protein breakdown rates. Compared with LG steers, HG steers had less initial BW but greater final BW, DMI (7.52 vs. 6.37 kg/d), ADG (1.33 vs. 0.853 kg/d), G:F (0.176 vs. 0.133 kg/kg), ME intake (0.233 vs. 0.201 Mcal x kg of BW(0.75) x d(-1)), and retained energy (RE; 0.0711 vs. 0.0558 Mcal x kg of BW(0.75) x d(-1)); gained more fat (676 vs. 475 g/d); and tended to gain more whole body protein (100 vs. 72 g/d), with no difference in residual feed intake (RFI). Estimated net energetic efficiency of gain (k(g)) and ME for maintenance (ME(m)) did not differ between the 2 groups, averaging 0.62 and 0.114, respectively. The HG steers had greater HCW (350 vs. 329 kg), backfat (16.1 vs. 11.6 mm), and yield grades (3.53 vs. 2.80), with a similar dressing percent, KPH fat, LM area, and marbling score. Skeletal muscle protein gain (70.2 vs. 57.6 g/d) and fractional protein accretion rate (0.242 vs. 0.197%/d) tended to be greater in HG than in LG steers. Steers were classified into low (-0.367 kg/d) and high (0.380 kg/d) RFI classes. Compared with the high RFI steers, low RFI steers consumed less DM (6.61 vs. 7.52 kg/d) and ME (0.206 vs. 0.234 Mcal x kg of BW(0.75) x d(-1)) and tended to gain less fat (494 vs. 719 g/d), but were similar for initial and final BW, ADG, G:F, protein gain, HCW, dressing percent, backfat, KPH fat, LM area, marbling score, and yield grade, as well as for all observations related to myofibrillar protein metabolism. Residual feed intake may be positively [corrected] correlated with ME for maintenance. The maintenance energy requirement increased by 0.0166 Mcal x kg(-0.75) x d(-1) for each percentage increase in fractional protein degradation rate, confirming the importance of this process in the energy economy of the animal.

Development of a Diagnostic Method for Detecting Increased Muscle Protein Degradation in Patients with Catabolic Conditions

Muscle atrophy in catabolic illnesses is due largely to accelerated protein degradation. Unfortunately, methods for detecting accelerated muscle proteolysis are cumbersome. The goal of this study was to develop a method for detecting muscle protein breakdown and assess the effectiveness of anticatabolic therapy. In rodent models of catabolic conditions, it was found that accelerated muscle protein degradation is triggered by activation of caspase-3. Caspase-3 cleaves actomyosin/myofibrils to form substrates for the ubiquitin-proteasome system and leaves a characteristic 14-kD actin fragment in the insoluble fraction of a muscle lysate. Muscle biopsies were obtained from normal adults and three groups of patients: 14 who were undergoing hip arthroplasty, 28 hemodialysis patients who were participating in exercise programs, and seven severely burned patients. In muscle of patients who were undergoing hip arthroplasty, the 14-kD actin fragment level was correlated (r = 0.787, P < 0.01) with the fractional rate of protein degradation. In muscle of hemodialysis patients who were undergoing endurance exercise training, the 14-kD actin fragment decreased to values similar to levels in normal adults; strength training did not significantly decrease the actin fragment. Severely burned patients had increased muscle protein degradation and actin fragment levels, but the two measures were not significantly correlated. The experimental results suggest that the 14-kD actin fragment in muscle biopsies is increased in catabolic states and could be used in conjunction with other methods to detect and monitor changes in muscle proteolysis that occur in patients with mild or sustained increases in muscle proteolysis.

GLP-2 stimulates intestinal growth in premature TPN-fed pigs by suppressing proteolysis and apoptosis.

We wished to determine whether exogenous glucagon-like peptide (GLP)-2 infusion stimulates intestinal growth in parenterally fed immature pigs. Piglets (106-108 days gestation) were given parenteral nutrient infusion (TPN), TPN + human GLP-2 (25 nmol. kg(-1). day(-1)), or sow's milk enterally (ENT) for 6 days. Intestinal protein synthesis was then measured in vivo after a bolus dose of [1-(13)C]phenylalanine, and degradation was calculated from the difference between protein accretion and synthesis. Crypt cell proliferation and apoptosis were measured in situ by 5-bromodeoxyuridine (BrdU) and terminal dUTP nick-end labeling (TUNEL), respectively. Intestinal protein and DNA accretion rates and villus heights were similar in GLP-2 and ENT pigs, and both were higher (P < 0.05) than in TPN pigs. GLP-2 decreased fractional protein degradation rate, whereas ENT increased fractional protein synthesis rate compared with TPN pigs. Percentage of TUNEL-positive cells in GLP-2 and ENT groups was 48 and 64% lower, respectively, than in TPN group (P < 0.05). However, ENT, but not GLP-2, increased percentage of BrdU-positive crypt cells above that in TPN piglets. We conclude that GLP-2 increases intestinal growth in premature, TPN-fed pigs by decreasing proteolysis and apoptosis, whereas enteral nutrition acts via increased protein synthesis and cell proliferation and decreased apoptosis.

Life‐history strategies and protein metabolism in overwintering juvenile Atlantic salmon: growth is enhanced in early migrants through lower protein turnover

The specific growth rate (length) of early migrant juvenile Atlantic salmon Salmo salar was significantly greater than that of delayed migrants in November and February. However, there were no differences in the fractional rate of protein synthesis (ks) of white muscle tissue between the migrant groups at any time. Early migrants had significantly greater fractional rates of protein growth (kg) of white muscle in February and significantly lower fractional rates of protein degradation (kd) in May. These results suggest that, compared to delayed migrants, Atlantic salmon adopting the strategy of early seaward migration maximize their overwinter growth by minimizing their rate of protein turnover.

Protein kinetics in callipyge lambs.

The objectives for this experiment were to determine the effect of the callipyge phenotype on protein kinetics. We studied callipyge and normal lambs (n = 37) at 5, 8, and 11 wk of age (n = 4 to 7/ group) to determine how protein kinetics are altered by this trait. Total protein, DNA, and RNA and calpastatin activity were measured in five skeletal muscles and in the heart, kidneys, and liver, and protein accretion rates were calculated. At 8 wk, the fractional synthesis rates of proteins in these tissues were measured in vivo using a primed, continuous 8-h infusion of [2H5]phenylalanine. Fractional rates of protein degradation were estimated by differences. At 5 wk of age, muscle weights, protein mass, protein:DNA, RNA:DNA, and calpastatin activity were higher (P < .05) for callipyge, and protein mass differences continued to increase through 11 wk. At 8 wk, fractional rates of protein synthesis and degradation were lower (P < .05) in callipyge than in normal lambs. The organs of callipyge lambs exhibited reduced growth at 11 wk. Thus, enhanced muscle growth seems to be maintained in callipyge lambs by reduced protein degradation rather than increased protein synthesis. However, we cannot exclude the possibility that the initial onset of the callipyge condition may be caused by an increase in the fractional rate of protein synthesis.

Mechanism for the increased skeletal muscle protein degradation in the obese zucker rat

Obese (fa/fa) Zucker rats showed a decreased protein content in skeletal muscle compared with their lean counterparts. This was associated with both a decrease in the fractional rate of protein synthesis and an enhanced fractional rate of protein degradation in skeletal muscle, as studied by pre-loading body proteins with 14C-bicarbonate. The increased protein degradation could be the result of the clearly enhanced expression for several transcripts of the ubiquitin genes in skeletal muscle. The results suggest that the increased protein degradation in the skeletal muscle of the obese animals may be due to increased activity of the ubiquitin-dependent proteolytic system.

Dexamethasone inhibits small intestinal growth via increased protein catabolism in neonatal pigs.

Our objective was to determine how dexamethasone (Dex) affects gastrointestinal protein metabolism and growth in neonatal pigs. Two-day-old pigs were given daily subcutaneous injections of either Dex (1 mg/kg body wt, n = 7) or saline (control, n = 6) for 7 days. In vivo protein synthesis was measured after 7 days with a bolus of [3H]phenylalanine. Tissue protein contents were measured in an initial control group of 2-day-old pigs and in control and Dex pigs after 7 days to estimate protein accretion and degradation. In control pigs, the protein accretion in the ileum was nearly sixfold greater than in the jejunum during the 7-day period. Dex nominally altered stomach growth but completely blocked the accretion of protein and DNA in the jejunum and ileum, with reduced villus height in the ileum. Dex increased the fractional protein degradation rate in the ileum (28%) and decreased the absolute protein synthesis rate in the jejunum and ileum by 17 and 21%, respectively. Dex resulted in a 40% lower total intestinal lactase activity compared with controls via reductions in both specific activity and tissue mass, especially in the ileum. Dex significantly decreased the circulating concentrations of insulin-like growth factor (IGF) I and IGF-binding protein (IGFBP)-1, -2, and -3. However, the tissue abundance of the IGF-I receptor in the stomach and ileum was greater in Dex pigs than controls. Our results suggest that Dex significantly inhibits small intestinal growth via both increased degradation and decreased synthesis of protein. Furthermore, the inhibition of intestinal growth resulted in significantly decreased lactose digestive capacity.

Selective changes in the protein-turnover rates and nature of growth induced in trout liver by long-term starvation followed by re-feeding.

We report upon the effects of a cycle of long-term starvation followed by re-feeding on the liver-protein turnover rates and nature of protein growth in the rainbow trout (Oncorhynchus mykiss). We determined the protein-turnover rate and its relationship with the nucleic-acid concentrations in the livers of juvenile trout starved for 70 days and then re-fed for 9 days. During starvation the total hepatic-protein and RNA contents decreased significantly and the absolute protein-synthesis rate (A(S)) also fell, whilst the fractional protein-synthesis rate (K(S)) remained unchanged and the fractional protein-degradation rate (K(D)) increased significantly. Total DNA content, an indicator of hyperplasia, and the protein:DNA ratio, an indicator of hypertrophy, both fell considerably. After re-feeding for 9 days the protein-accumulation rates (K(G), A(G)) rose sharply, as did K(S), A(S), K(D)), protein-synthesis efficiency (K(RNA)) and the protein-synthesis rate/DNA unit (K(DNA)). The total hepatic protein and RNA contents increased but still remained below the control values. The protein:DNA and RNA:DNA ratios increased significantly compared to starved fish. These changes demonstrate the high response capacity of the protein-turnover rates in trout liver upon re-feeding after long-term starvation. Upon re-feeding hypertrophic growth increased considerably whilst hyperplasia remained at starvation levels.

Protein turnover in skeletal muscle of tumour-bearing transgenic mice overexpressing the soluble TNF receptor-1

The implantation of the Lewis lung carcinoma (a fast-growing mouse tumour that induces cachexia) to both wild-type and transgenic mice for the soluble TNF receptor type I protein (sTNF-R1) resulted in a considerable loss of carcass weight in both groups. However, while in the wild-type mice there was a loss of both fat and muscle, in the transgenic mice muscle waste was not affected to the same extent as in the wild-type group. Muscle waste in wild-type mice was accompanied by an increase in the fractional rate of protein degradation, while no changes were observed in protein synthesis. The result was a decreased rate of protein accumulation which accounted for the muscle weight loss observed as a result of the tumour burden. In contrast, transgenic mice did not have such low rates of protein accumulation after tumour implantation. The increase in protein degradation in the tumour-bearing transgenic mice was accompanied by a similar increase in protein synthesis which compensated for the loss of muscle protein by degradation. Both tumour-bearing groups showed an enhanced expression of ubiquitin and proteasome C8 subunit genes, all of them related to the activation of the ATP-dependent proteolytic system in skeletal muscle. It is suggested that TNF may, in part, be responsible for the loss of protein in skeletal muscle of tumour-bearing mice.

Mechanisms of cardiac hypertrophy in canine volume overload.

This study tested whether the modest hypertrophy that develops in dogs in response to mitral regurgitation is due to a relatively small change in the rate of protein synthesis or, alternatively, is due to a decreased rate of protein degradation. After 3 mo of severe experimental mitral regurgitation, the left ventricular (LV) mass-to-body weight ratio increased by 23% compared with baseline values. This increase in LV mass occurred with a small, but not statistically significant, increase in the fractional rate of myosin heavy chain (MHC) synthesis (Ks), as measured using continuous infusion with [3H]leucine in dogs at 2 wk, 4 wk, and 3 mo after creation of severe mitral regurgitation. Translational efficiency was unaffected by mitral regurgitation as measured by the distribution of MHC mRNA in polysome gradients. Furthermore, there was no detectable increase in translational capacity as measured by either total RNA content or the rate of ribosome formation. These data indicate that translational mechanisms that accelerate the rate of cardiac protein synthesis are not responsive to the stimulus of mitral regurgitation. Most of the growth after mitral regurgitation was accounted for by a decrease in the fractional rate of protein degradation, calculated by subtracting fractional rates of protein accumulation at each time point from the corresponding Ks values. We conclude that 1) volume overload produced by severe mitral regurgitation does not trigger substantial increases in the rate of protein synthesis and 2) the modest increase in LV mass results primarily from a decrease in the rate of protein degradation.

Role of TNF receptor 1 in protein turnover during cancer cachexia using gene knockout mice

The implantation of the Lewis lung carcinoma (a fast-growing mouse tumour that induces cachexia) to both wild-type and gene-deficient mice for the TNF- α receptor type I protein (Tnfr1°/Tnfr1°), resulted in a considerable loss of carcass weight in both groups. However, while in the wild-type mice there was a loss of both fat and muscle, in the gene-knockout mice muscle wastage was not affected to the same extent. In both groups, tumour burden resulted in significant increases in circulating TNF- α, a cytokine which, as we have previously demonstrated, can induce protein breakdown in skeletal muscle. Muscle wastage in wild-type mice was accompanied by an increase in the fractional rate of protein degradation, while no changes were observed in protein synthesis. The result is a decreased rate of protein accumulation that accounts for the muscle weight loss observed as a result of tumour burden. In contrast, gene knockout mice did not have significantly lower rates of protein accumulation as a result of tumour implantation. The increase in protein degradation in the tumour-bearing wild mice was accompanied by an enhanced expression of both ubiquitin and proteasome subunit genes, all of them related to the activation of the ATP-dependent proteolytic system in skeletal muscle. Tumour-bearing gene-deficient mice did not show any increase in gene expression. It is concluded that TNF- α (alone or in combination with other cytokines) is responsible for the activation of protein breakdown in skeletal muscle of tumour-bearing mice.

In vivo unaltered muscle protein synthesis in experimental chronic metabolic acidosis

Chronic metabolic acidosis (CMA) is a major cause of growth defect, implying disturbances of protein metabolism. Previously, in vivo studies performed in the fasting state showed enhanced whole body protein turnover, whereas in vitro studies showed unchanged muscle protein synthesis. The present study is the first to determine the effects of CMA on muscle protein synthesis and degradation in vivo. Two studies were performed in 60 g male rats fed a 30% casein diet. In study I, one group was sham-operated (C rats), and two groups underwent subtotal nephrectomy. One of them developed acidosis (UA rats) which was corrected in the other by NaHCO3 in the diet (UNA rats). Study II compared sham-operated rats rendered acidotic by NH4Cl in the drinking water (CA rats) and normal pair-fed (CNA) rats. Fractional protein synthesis rate (FSR) was determined in gastrocnemius muscle after injection of 3H-phenylalanine. Fractional protein degradation rate (FDR) was calculated as FSR minus fractional rate of muscle growth (FGR). In study I, UA rats had lower growth and N balance (163 +/- 12 vs. 216 +/- 11 mg N/day; P < 0.001) than UNA rats, despite identical food intake (11 g/day). This was associated with identical FSR (10.4 +/- 0.5 vs. 10.9 +/- 0.5%/day), but enhanced protein degradation (6.30 +/- 0.99 vs. 5.10 +/- 0.71%/day; P < 0.05). Plasma insulin, C peptide, PTH and corticosterone did not differ in UA and UNA rats, whereas plasma IGF-I was markedly reduced (147 +/- 21 vs. 283 +/- 27 ng/ml; P < 0.01) in UA rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Tumor necrosis factor-alpha mediates changes in tissue protein turnover in a rat cancer cachexia model.

Rats bearing the Yoshida AH-130 ascites hepatoma showed enhanced fractional rates of protein degradation in gastrocnemius muscle, heart, and liver, while fractional synthesis rates were similar to those in non-tumor bearing rats. This hypercatabolic pattern was associated with marked perturbations of the hormonal homeostasis and presence of tumor necrosis factor in the circulation. The daily administration of a goat anti-murine TNF IgG to tumor-bearing rats decreased protein degradation rates in skeletal muscle, heart, and liver as compared with tumor-bearing rats receiving a nonimmune goat IgG. The anti-TNF treatment was also effective in attenuating early perturbations in insulin and corticosterone homeostasis. Although these results suggest that tumor necrosis factor plays a significant role in mediating the changes in protein turnover and hormone levels elicited by tumor growth, the inability of such treatment to prevent a reduction in body weight implies that other mediators or tumor-related events were also involved.

The degradability of legume seed meals in the rumen: effect of heat treatment

The extent and rate of degradation of dry matter and nitrogen in ground seeds of pea ( Pisum sativum), lupin ( Lupinus albus var. multolupa), field bean ( Vicia faba var. minor), vetch ( Vicia sativa) and bitter vetch ( Vicia ervilia), previously autoclaved (A) or not (NA) at 120 °C for 30 min, was measured using nylon bags incubated in the rumen of wethers fed lucerne hay. The fractional outflow rate of the legume seed meal particles was determined with chromium as a marker. More than 88% of the protein in untreated legume seed meals, except for vetch, disappeared from the bags by 24 h. Autoclaving decreased both the soluble fraction and the fractional rate of protein degradation of the slowly degraded fraction ( P < 0.01). Rumen outflow rate was, respectively, 0.0154 h −1 and 0.0222 h −1, at 0.7 times and 1.0 times maintenance level. Effective degradability of protein was strongly reduced by heat treatment in pea and lupin seeds, moderately decreased in field bean and bitter vetch and almost unchanged in vetch seed meal.

Whole-body and tissue protein synthesis during brief and prolonged fasting in the rat

1. Little information is currently available on protein turnover during chronic protein loss situations. We have thus measured the whole-body and tissue protein fractional synthesis rates (ks), the whole-body fractional protein degradation rate (kd), the capacity for protein synthesis (Cs) and the efficiency of protein synthesis (kRNA) in vivo in fed and fasted (1, 5 and about 9 days) 400 g rats. 2. One day of starvation resulted in a reduced ks and an increased kd in the whole body. ks was selectively depressed in skeletal muscles, mainly owing to a reduced kRNA, and was not modified in heart, liver and skin. The contribution of skin to whole-body protein synthesis increased by 39%. 3. During the phase of protein sparing (5 days of fasting), kd in the whole body decreased below the control fed level. ks in skeletal muscles was sustained because kRNA was restored to 82-98% of the control value. 4. Rats were in a protein-wasting phase after 9 days of starvation. kd in the whole body did not increase and was actually 78% of the value observed in fed animals. By contrast, ks in the whole body and tissues decreased to 14-34% of the control values, owing to reductions in both Cs and kRNA. Whatever the duration of the fast, the contribution of the skin to whole-body protein synthesis largely exceeded that of skeletal muscle. 5. The present findings suggest that the main goal in the treatment of chronic protein loss should be to sustain protein synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

Activity measurement of lysosomal cysteine proteinases, cathepsins B, H and L, in crude tissue extracts, and their relation to the fractional rate of protein degradation

1. 1. The fractional rate of protein degradation was measured in vivo and compared with the lysosomal enzyme activities of several rat tissues. Good agreement was observed between them. 2. 2. A specific method for determining lysosomal cysteine-dependent cathepsins, B, L and H, was demonstrated.