Quantitative determination of urinary N-tau-methylhistidine output as an index of myofibrillar protein degradation

N tau-Methylhistidine(3-methylhistidine) in urine of the rat is mainly derived from the degradation of actin and myosin in skeletal muscle, intestine and skin. The fractional degradation rates of the myosin-actin pools of these tissues were calculated from the time course of increase in the specific radioactivities of N tau-methylhistidine after daily administration of [methyl-14C]methionine to young adult rats under conditions of restricted food intake. The contributions to urinary excretion of N tau-methylhistidine from the three tissues were calculated from the fractional degradation rates and N tau-methylhistidine contents of the three tissues; 75.6% for skeletal muscle, 2.2% for intestine and 22.2% for skin. The results show that the skeletal muscle is the major source of urinary N-tau-methylhistidine output, but the contribution of skin is not negligible in rats. The specific radioactivity of N tau-methylhistidine in urine was much higher than that of skeletal muscle. The fractional degradation rates of myosin and actin in skeletal muscle had similar values. Although the specific radioactivities of N tau-methylhistidine in myosin and actin were very different, the mean value was similar to that in mixed skeletal muscle.     

Response of urinary hydroxyproline to dietary protein and fasting in white-tailed deer

The effects of dietary protein, fasting, and refeeding on urinary hydroxyproline of nine captive female white-tailed deer (Odocoileus virginianus) were examined from 23 February to 3 May 1984 in northern Minnesota. In the fasted group, mean hydroxyproline:creatinine (OHP:C) was greater (P less than 0.05) at week 4 compared to baseline at week 0. Between fasted deer and deer fed high protein-high energy (HPHE) and low protein-high energy (LPHE) diets, no difference in OHP:C ratios was detected during the initial 4 wk of the study. Urinary OHP:C ratios were significantly (P less than 0.05) greater in the fasted group during refeeding, concomitant with greater feed consumption and weight gain. There was also a significant (P less than 0.02) time effect in the fasted-refed group; OHP:C ratios increased during these two phases of the study. There was no difference between the HPHE and LPHE fed deer in renal OHP excretion. However, mean OHP:C ratios in yearlings (16.8 +/- 2.2) were greater (P less than 0.001) than in the adults (7.5 +/- 1.2) of those groups, indicating a higher collagen turnover rate. Urinary OHP:C shows potential as an indicator of growth and starvation, and the data presented may serve as reference values.     

Response of muscle protein turnover to insulin after acute exercise and training.

To determine whether the enhanced insulin-sensitivity of glucose metabolism in muscle after acute exercise also extends to protein metabolism, untrained and exercise-trained rats were subjected to an acute bout of exercise, and the responses of protein synthesis and degradation to insulin were measured in epitrochlearis muscles in vitro. Acute exercise of both untrained and trained rats decreased protein synthesis in muscle in the absence or presence of insulin, but protein degradation was not altered. Exercise training alone had no effect on protein synthesis or degradation in muscle in the absence or presence of insulin. Acute exercise or training alone enhanced the sensitivities of both protein synthesis and degradation to insulin, but the enhanced insulin-sensitivities from training alone were not additive to those after acute exercise. These results indicate that: a decrease in protein synthesis is the primary change in muscle protein turnover after acute exercise and is not altered by prior exercise training, and the enhanced insulin-sensitivities of metabolism of both glucose and protein after either acute exercise or training suggest post-binding receptor events.     

Response of storage protein levels to variation in dietary protein levels

Storage proteins have been found to play a major role in insect metamorphosis and egg production and are accumulated during the actively feeding larval stage. Yet few studies have focused on how nutrition affects storage protein levels. Three storage proteins were identified in male and female Heliothis virescens pupae, one arylphorin and two putative high-methionine hexamers. Storage proteins were quantified in early pupae and in pharate adults. Storage protein levels peaked in 48-h pupae and were more abundant in females across all stages. Both male and female pharate adults retained a portion of total storage protein levels and females retained greater levels overall. In females, post-eclosion protein reserves will likely be used toward egg manufacturing, while the role of protein reserves in males remains speculative. In our previous study of H. virescens larvae, we found that protein-derived growth in females progressively increased as dietary protein levels increased. Our present data show that levels of storage protein also increased progressively along with dietary protein levels. This suggests that females allocated protein, in excess of adult tissue formation needs, toward storage protein. Our study is the first to demonstrate how responsive storage protein levels can be in face of varying levels of dietary protein.     

Effect of dietary lysine on muscle protein turnover in growing chickens.

Day-old male chickens were fed ad libitum isoenergetic diets containing 20% crude protein but differing in their lysine content (from 6.5 up to 11.3 g/kg). At 3 weeks of age, protein fractional synthesis rates in the pectoralis major muscle were determined using a large dose injection of 120 mumol per kg body weight of L-[4-3H] phenylalanine. Protein gain in the pectoralis major was measured between 19 and 23 days of age. Protein breakdown was obtained by calculating the difference between protein synthesis and deposition. Weight gain varied curvilinearly with dietary lysine intake and was maximum for 11.3 g lysine/kg of diet. In birds fed an adequate lysine intake (10.1-11.3 g/kg) protein fractional synthesis and breakdown rates were 23.6-25.9 and 17.8-19.8%/d respectively. Increasing lysine supplementation in the diet resulted in an impairment of protein fractional breakdown rates. By contrast, protein fractional synthesis rates remained unchanged owing mainly to an improvement in the synthesis efficiency (kRNA), until birds were fed an adequate lysine intake. These data suggest that the growth rate reduction of chickens fed lysine deficient diets was due to alterations in both rates of protein synthesis and breakdown in skeletal muscle. A maximum protein deposition is achieved when kRNA was optimal, ie for a dietary lysine content of about 9 g/kg, a value close to the requirement.     

Effect of dietary protein and tryptophan and the turnover of rat liver ornithine aminotransferase.

Under controlled conditions of diet, age, and animal management we confirmed our earlier studies that the fractional turnover rate constant of rat liver ornithine aminotransferase (L-ornithine:2-oxoacid aminotransferase, EC 2.6.1.13) was about 0.4 day-1 when estimated by tracer technique. However, when estimated by the kinetics of enzyme activity perturbation during dietary induction or return to normal levels, a value of about 0.7 day-1 was obtained. This discrepancy may now be attributed in part to a transistory change in the fractional rate constant during enzyme adaption.     

Myofibrillar protein turnover. Synthesis rates of myofibrillar and sarcoplasmic protein fractions in different muscles and the changes observed during postnatal development and in response to feeding and starvation.

Measurement of rates of synthesis of skeletal-muscle proteins in adult rats shows that the faster overall rate of turnover in diaphragm and soleus muscles compared with several other, more glycolytic, muscles is also exhibited by the myofibrillar proteins, since the ratio of sarcoplasmic to myofibrillar protein synthesis is similar for all muscles. Further, throughout postnatal development, when the overall turnover rate falls with age, parallel changes occur for the myofibrillar proteins, as indicated by a constant ratio of sarcoplasmic to myofibrillar protein synthesis (2.06) in the steady state after overnight starvation. Only in the youngest (4 weeks old) rats is a slightly lower ratio observed (1.72). These results indicate that, when changes in the overall turnover rate of muscle proteins occur, the relative turnover of the two major protein fractions stays constant. However, measurements in the non-steady state during growth and after starvation for 4 days show that the relative synthesis rates of the two fractions change as a result of a disproportionate increase in myofibrillar protein synthesis during growth and decrease during starvation. Thus the synthesis rate of the slower-turning-over myofibrillar protein fraction is more sensitive to nutritional state than is that of the sarcoplasmic protein. It is suggested that such responses may help to maintain constant tissue composition during non-steady-state conditions of growth and atrophy.     

Skeletal-muscle growth and protein turnover.

Because of turnover, protein synthesis and breakdown can each be involved in the regulation of the growth of tissue protein. To investigate the regulation of skeletal-muscle-protein growth we measured rates of protein synthesis and breakdown in growing rats during development on a good diet, during development on a marginally low-protein diet and during rehabilitation on a good diet after a period of severe protein deficiency. Rates of protein synthesis were measured in vivo with a constant intravenous infusion of [14C]tyrosine. The growth rate of muscle protein was measured and the rate of breakdown calculated as breakdown rate=synthesis rate-growth rate. These measurements showed that during development on a good diet there was a fall with age in the rate of protein synthesis resulting from a fall in capacity (RNA concentration) and activity (synthesis rate per unit of RNA). There was a fall with age in the breakdown rate so that the rate was highest in the weaning rats, with a half-life of 3 days. There was a direct correlation between the fractional growth and breakdown rates. During rehabilitation on the good diet, rapid growth was also accompanied by high rates of protein breakdown. During growth on the inadequate diet protein synthesis rates were lesss than in controls, but growth occurred because of decreased rates of protein breakdown. This compression was not complete, however, since ultimate muscle size was only one-half that of controls. It is suggested that increased rates of protein breakdown are a necessary accompaniment to muscle growth and may result from the way in which myofibrils proliferate.     

Distinctive features of dietary phosphate supply.

Dietary phosphate has profound effects on growth and renal handling of the compound. On the basis of changes in growth rate and food intake, after alterations in phosphate load, our laboratory previously suggested that these effects are mediated by intestinal signals (Landsman A, Lichtstein D, Bacaner M, and Ilani A. Br J Nutr 86: 217-223, 2001). The aim of this study was to further evaluate the role of dietary phosphate on food intake and appetite and specific organ growth, and to test for the presence of a serum factor that may affect renal phosphate handling in phosphate-resupplied rats. The experimental design was based on a comparison between groups of rats receiving identical low-phosphate diets but drinking water containing either phosphate or chloride. We show that 1) changes in food intake after alterations in phosphate load occurred in parallel with variations in digestive system distention, suggesting that dietary phosphate has also a direct effect on appetite; 2) dietary phosphate-dependent growth has a specific effect on the growth of liver and epididymal fat; and 3) serum of rats supplied with phosphate contains a factor that inhibits sodium-dependent phosphate transport in a model of renal proximal tubule cells. Collectively, these observations are in accord with the hypothesis that factor(s) emanating from the digestive system in response to dietary phosphate load may be involved in growth, appetite and renal handling of phosphate.     

Information-theoretic significance of Gibbs energy supply to editing mechanisms.

If a branched multistep editing mechanism is implemented by an enzyme with a single site for the specific substrates, there is no reason to believe that the number of testing steps is fixed and cannot be controlled by some external factors. The paper considers the mechanisms of single- or multistep editing done in response to various factors, particularly to the value of displacement from the reaction equilibrium. To avoid a complicated analysis of a fully specified case, which would be likely to obscure the general significant features, the operating modes of those mechanisms are estimated using the method of minimizing associated information gains. It turns out that sufficiently far from equilibrium the variable mechanisms can essentially operate just as well as the fixed multistep mechanisms.     

Myofibrillar protein oxidation and contractile dysfunction in hyperthyroid rat diaphragm.

The purpose of the present study was to test the hypothesis that administration of thyroid hormone [3,5,3'-triiodo-L-thyronine (T(3))] could result in oxidation of myofibrillar proteins and, in turn, induce alterations in respiratory muscle function. Daily injection of T(3) for 21 days depressed isometric forces of diaphragm fiber bundles across a range of stimulus frequencies (1, 10, 20, 40, 75, and 100 Hz) (P < 0.05). These reductions in force production were accompanied by a remarkable increment (104%; P < 0.05) in carbonyl groups of myofibrillar proteins. In contrast, T(3) treatment has no effects on the carbonyl content in myosin heavy chain. In additional experiments, we have also tested the efficacy of carvedilol, a nonselective beta(1)- beta(2)-blocker that possesses antioxidative properties. Treatment with carvedilol dramatically improved isometric tetanic force production at stimulus frequencies from 40 to 100 Hz (P < 0.05). Carvedilol also prevented T(3)-induced contractile protein oxidation (P < 0.05). These data suggest that the oxidative modification of myofibrillar proteins may account, at least in part, for an impairment of diaphragm in hyperthyroidism.