Recent advances in elucidating the mechanisms that control body protein synthesis and degradation both expand and complicate our understanding of how these processes are regulated. This review presents an introduction to the multiple regulatory systems involved, emphasizing the number of potential controls. These include gene transcription, gene activation or suppression, activation or suppression of mRNA translation and activation or suppression of signaling pathways. The complexity of these interacting controls presents a challenge to our understanding of the overall coordinated regulation of protein synthesis and degradation and its response to any particular stimulus. Specific examples are used to illustrate regulatory mechanisms, including the ways in which protein metabolism is regulated by the amino acid leucine. In addition to regulation associated with gene expression and post-translational control, the expanding field of epigenetics adds another layer of complexity, including trans-generational responses to nutrient intake, highlighting the potential for long-term impact of nutritional experience on the metabolism of subsequent generations.

This study was carried out to determine ruminal dry matter (DM) and crude protein (CP) degradation characteristics of untreated, 2-, 4- and 6-min microwave-treated soya-bean meal (SBM) by using nylon bags and sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE) techniques. Nylon bags of untreated or treated SBM were suspended into the rumen of three Holstein steers from 0 to 48 h, and data were fitted to non-linear degradation characteristics to calculate effective rumen degradation (ERD). There were significant differences (P < 0·05) for DM and CP degradation parameters between untreated and microwave-treated SBM. Microwave treatments decreased the water-soluble fraction and increased the potentially degradable fraction of CP. The degradation rate of the latter fraction decreased with these treatments. As a consequence, microwave treatments decreased (P < 0·05) ERD of CP. From densitometric scanning, SBM proteins were seen to be composed of two major components; β-conglycinin and glycinin, accounting for proportionately 0·30 and 0·40 of buffer-soluble SBM proteins, respectively. Electrophoretic analysis of untreated, 2-, 4- and 6-min microwave-treated SBM protein residues revealed that two of the subunits of β-conglycinin (α -and α) were degraded completely after 2, 4, 24 and 48 h, respectively, whereas the α subunit of this protein was more resistant to degradation. In untreated SBM, the two subunits of glycinin (acidic and basic polypeptides) were degraded in the middle of the incubation period, but in microwave-treated SBM were not degraded until 48 h of incubation. In vitro digestibility of ruminally undegraded CP of untreated and treated SBM increased (P < 0·05) with increases in rumen incubation time from 8 to 24 h. In conclusion, SBM proteins appeared to be effectively protected from ruminal degradation by a 4-min microwave treatment. SDS-PAGE results indicated that ruminally undegraded protein from untreated SBM was mainly composed of the basic subunit of glycinin, whereas that from microwave-treated SBM was composed of β-conglycinin and both basic and acidic subunits of glycinin.

Diets with protein of inferior quality may increase protein breakdown in skeletal muscle but the experimental results are inconsistent. To elucidate the relationship, pigs were fed isoenergetic and isonitrogenous diets based on soyabean-protein isolate or casein for 15 weeks, with four to six animals per group. A higher plasma level of urea (2.5-fold the casein group value, P=0.01), higher urinary N excretion (2.1-fold the casein group value, P=0.01), a postabsorptive rise in the plasma levels of urea, 3-methylhistidine and isoleucine in soyabean protein-fed pigs suggested recruitment of circulatory amino acids by protein breakdown in peripheral tissues. Significant differences between dietary groups were detected in lysosomal and ATP-dependent proteolytic activities in the semimembranosus muscle of food-deprived pigs. A higher concentration of cathepsin B protein was found, corresponding to a rise in the cathepsin B activity, in response to dietary soyabean protein. Muscle ATP-stimulated proteolytical activity was 1.6-fold the casein group value (P=0.03). A transient rise in the level of cortisol (2.9-times the casein group value, P=0.02) occurred in the postprandial phase only in the soyabean group. These data suggest that the inferior quality of dietary soyabean protein induces hormonally-mediated upregulation of muscle protein breakdown for recruitment of circulatory amino acids in a postabsorptive state.

Both the peptidase activity against small fluorescent peptide substrates and the ATPase activity of Lon (La) proteases are stimulated by unstructured proteins such as α-casein. This stimulation reveals the simultaneous interaction of Lon with two proteolytic substrates—α-casein and the peptide substrate. To understand the cellular function of this stimulation, it is important to determine the physical properties of Lon stimulators. The abilities of compositionally simple random copolymers of amino acids (rcAAs) to stimulate the peptidase and ATPase activities of the Lon protease from Mycobacterium smegmatis (Ms-Lon) and its N-terminal truncation mutant (N-E226) were determined. We report that cationic but not anionic rcAAs stimulated Ms-Lon's peptidase activity but were themselves poor substrates for the enzyme. Peptidase stimulation by rcAAs correlated approximately with the degree of hydrophobicity of these polypeptides and reached levels >10-fold higher than observed previously for Ms-Lon stimulators such as α-casein. In contrast to α-casein, which stimulates Ms-Lon's peptidase activity by 40% and ATPase activity by 150%, rcAAs stimulated peptidase activity without concomitant stimulation of ATPase activity. Active site labeling experiments suggested that both rcAAs and ATP increased peptidase activity by increasing accessibility to the peptidase active site. Peptidase activity assays in the presence of both α-casein and rcAAs revealed that interactions of rcAAs and α-casein with Ms-Lon are extremely complex and not mutually exclusive. Specifically, (1) additions of low concentrations of α-casein (<50 μg/mL) caused a further stimulation of Ms-Lon's rcAA-stimulated peptidase activity; (2) additions of higher concentrations of α-casein inhibited Ms-Lon's rcAA-stimulated peptidase activity; (3) additions of all concentrations of α-casein inhibited N-E226's rcAA-stimulated peptidase activity. We conclude the Ms-Lon can interact with an rcAA, α-casein, and a substrate peptide simultaneously, and that formation of this quaternary complex requires the N-terminal domain of Ms-Lon. These data support models of Ms-Lon that include two allosteric polypeptide binding sites distinct from the catalytic peptidase site.

The present study was undertaken to investigate the changes in muscle protein turnover involved in the rapid mobilization of protein in rats subjected to severe protein restriction during lactation. Estimates of mammary gland and liver protein synthesis were also made during lactation. Multiparous female Sprague-Dawley rats, caged individually following mating, were offered a high-protein diet (H; 215 g crude protein (N - 6·25; CP)/kg dry matter (DM)) ad lib. until parturition. Following parturition, half the females continued to receive diet H, whilst the remainder were offered a diet low in protein (L; 90 g CP/kg DM) ad lib. On days 2, 4, 8 and 12 of lactation, groups of females were used in the estimation of tissue protein synthesis (flooding dose of [3H]phenylalanine) immediately after a milk sample had been obtained. Rates of muscle protein synthesis were unchanged during lactation in group H. The feeding of diet L during lactation reduced the muscle protein synthesis on day 12 to rates that were lower than group H and also the rate on diet L on day 2 (P 0·01). However, this fall in muscle protein synthesis was not rapid and muscle fractional synthesis rate (FSR) was different from group H only from day 8 (P 0·05). Estimated rates of mammary protein synthesis appeared to be generally unchanged by dietary treatment or stage of lactation. Liver FSR was also unchanged by dietary protein supply or stage of lactation. The effect of dietary protein restriction on liver size and protein content during lactation influenced liver absolute synthesis rate (ASR), and on days 8 and 12 of lactation liver ASR was lower in group L than in group H (P 0·001). The loss of muscle protein in rats fed on diet L during lactation (133 mg) occurred mainly between days 2 and 8 of lactation and was primarily associated with a dramatic increase in degradation (13·0% per d), with the decline in synthesis having a much smaller role. A decline in muscle protein degradation during the latter half of lactation was part of the mechanism that prevented excessive muscle protein catabolism. It is thought that the estimation of mammary protein synthesis in the present study was impaired by the milk sampling procedure previously used.

Live-weight gain responses were investigated using 36 individually penned lambs (mean live weight 35·2 kg) given three sodium hydroxide treated straw-based diets: low-protein, low-rapeseed meal (control) diet; high protein, high-rapeseed meal (HR) diet; or high-protein, fish meal (FM) diet. The diets were formulated to provide 3 or 9 g undegradable nitrogen per kg dry matter (DM) respectively for the diets without or with fish meal. Diets were offered once a day in a 50: 50 forage-to-concentrate ratio in amounts calculated to support maintenance plus 150 g gain and were adjusted weekly according to live weight. Live-weight gain, measured for 7 weeks, was improved by the FM diet only (P < 0·05).

The three diets were given also to rumen-fistulated sheep. The FM diet maintained higher rumen ammonia concentrations during most of the day. The FM and HR diets reduced rumen solid particle outflow rate (P < 0·05) and increased the effective degradability of DM and acid-detergent fibre.

Urinary excretion of acid-soluble peptides (ASP) was measured in rats given streptozotocin or dexamethasone. Streptozotocin-induced diabetic rats excreted increased amounts of urinary nitrogen and ASP-form amino acids. The urinary ratio for N: ASP-form leucine plus valine, which has been shown to reflect the efficiency of dietary N utilization, increased in the diabetic rats, suggesting the impaired utilization of dietary N (and re-utilization of endogenous N). Dexamethasone administration to adrenalectomized rats caused increased excretion of urinary ASP-form leucine plus valine with a concomitant increase in N excretion. However, urinary ratio for N: ASP-form leucine plus valine did not change significantly. The results suggested that dexamethasone caused increased degradation of body proteins, but maintained the efficiency of dietary N utilization or re-utilization of endogenous N. Based on the present observations and the hypothesis proposed previously by the present authors (Noguchi et al. 1988) that urinary excretion of ASP-form leucine plus valine reflects the rate of whole-body protein degradation and the urinary ratio for N: ASP-form leucine plus valine represents N utilization, the origin of urinary N in streptozotocin-diabetic and dexamethasone-administered rats is discussed

Four concentrate supplements differing in crude protein (CP) and undegradable protein (UDP) content were offered to 16 lactating Friesian cows together with grass silage (dry matter (DM) 196 g/kg, pH 4·38, CP 160 g/kg DM, in vitro DM digestibility 0·68) ad libitum in a Latin-square trial with 3-week periods. The supplement treatments were: (1) barley 122 g CP per kg DM, degradability (dg) 0·77; (2) barley/soya-bean meal 210 g CP per kg DM, dg 0·69; (3) barley/soya-bean meal/fish meal 190 g CP per kg DM, dg 0·61; (4) barley/soya-bean meal/fish meal 219 g CP per kg DM, dg 0·59. Supplements were given at 8 kg/day. Total daily intakes of silage (kg DM), CP and UDP (g) on treatments 1 to 4 were 7·77, 2087, 375; 8·35, 2804, 655; 8·29, 2676, 717; 8·70, 2917, 826, respectively. Milk yield (kg/day) and yields of fat, protein and lactose (g/day) on the four treatments were 21·3, 791, 617, 984; 23·0, 816, 688, 1055; 23·0, 818, 696, 1050; 23·6, 813, 735, 1071 for treatments 1 to 4 respectively. Yield and concentration of protein and lactose were significantly lower on treatment 1 than on the other treatments, while the of blood metabolites indicated treatment effects on blood glucose, non-esterified fatty acids, plasma protein and urea. Digestibility of organic matter and non-ammonia nitrogen (NAN) flow to the abomasum (g/day), measured in sheep given a fixed silage/supplement ratio at maintenance, were 0·81 and 18·4, 0·81 and 20·8, 0·82 and 21·4, 0·82 and 22·4 for treatments 1 to 4 respectively. The NAN flow was significantly greater on treatment 4 than on treatment 1.

Crude protein concentration in diets based on high-moisture maize grain was increased using rumen degradable protein (RDP) or rumen undegradable protein (UDP). Treatment diets (numbers in parentheses indicate approximate RDP and UDP concentrations, g/kg dry matter (DM)) were: low-RDP (59, 80), high-RDP (79, 92), low-UDP (66, 67), high-UDP (69, 107, with maize gluten meal as chief UDP source) and high-UDP (71, 91, with fish, meat and maize gluten meals as UDP sources). Rumen pH and concentrations of volatile fatty acids, measured hourly for 12 h after feeding in four rumen fistulated Holstein steers, were unaffected by treatment (P > 0·05). Rumen ammonia-N concentration was greater for high-RDP (193 mg/1) than low-RDP (68 mg/1; P < 0·001). Within 12 h of feeding, ammonia-N concentration declined from 169 to 58 mg/1 for low-RDP and remained stable for high-RDP.

In a 56-day feeding trial using Holstein heifers (live weight 128 kg) and the first four treatments, RDP concentration had no effect on DM intake (P > 0·05). Lower DM intake (P < 0·01) for high-UDP (4·40 kg/day) than low-UDP (5·13 kg/day) was attributed to low palatability of maize gluten meal. Live-weight gain and food conversion efficiency were unaffected by treatment.

In a 56-day feeding trial using Holstein bulls (live weight 110 kg) and all treatments, RDP concentration had no effect on animal performance (P > 0·05). Food conversion efficiency increased (P < 0·001) when UDP concentration was increased only if UDP was supplied by a mix of fish, meat and maize gluten meals.

When RDP supply was not limiting to DM intake, protein source (amino acid composition) was better than protein degradability as an index of the feeding value of dietary protein.