The changing pattern of whole body protein metabolism in aging humans.

Dynamic aspects of whole body protein (nitrogen) metabolism were explored in healthy young adults and elderly men and women. Measurements were made of the rate of whole body protein breakdown, with the aid of 15N-glycine, and the rate of muscle protein breakdown, as estimated from urinary N tau-methylhistidine excretion. The results also were evaluated in relation to obligatory (endogenous) urinary nitrogen losses, previously determined in this laboratory for the two age groups. Rates of whole body and muscle protein breakdown, per unit body weight, were lower in elderly subjects than in young adults. Muscle accounted for a mean of 27% of whole body protein breakdown in young adults and 20% or less (p less than 0.01) in elderly subjects. Daily obligatory N loss was positively correlated (p less than 0.01) with whole body protein breakdown. It was calculated that muscle contributed less to the obligatory N output in elderly subjects than in young adults. These results indicate a change in the distribution of whole body protein metabolism during aging in human subjects, with muscle making a lower contribution to total body protein metabolism in elderly subjects compared with young adults.

Musle protein breakdown rates in humans based on Ntau-methylhistidine (3-methylhistidine) content of mixed proteins in skeletal muscle and urinary output of Ntau-methylhistidine.

Samples of psoas muscle from nine infants (aged 1 day to 14 mo) and of several skeletal muscles from seven adult males (age 19-74 yr) were analyzed for content of protein-bound Ntau-methylhistidine (3-methylhistidine; 3-Mehis). The mean content of 3-Mehis (expressed as mumoles/g mixed protein) was 3.2 (range 2.4-3.7) in infants and 4.2 (range 3.7-4.6) in adults. The daily urinary excretion of 3-Mehis was measured in four young adult males receiving an egg-protein, flesh-free diet. Mean excretion of 3-Mehis was 211 (range 167-252) mumoles/day. From these two sets of data the mean rate of muscle protein breakdown in adult males was estimated to be 50 g/day, or 0.7 +/- 0.1 g/kg body weight/day. These results are compared with reported values for the 3-Mehis content of mixed proteins in muscle of various species, and with published estimates, computed by other techniques, of the rate of muscle protein breakdown in human subjects.

Myofibrillar protein turnover and urinary N-tau-methylhistidine output. Response to dietary supply of protein and energy.

The urinary excretion of total N-tau-methylhistidine by the growing rat was measured to evaluate the effects of dietary protein and energy restriction on muscle protein turnover in vivo. 2. Young male rats (about 100 g initial wt.) were fed on one of three diets. Group I (controls) received an adequate 18% lactalbumin diet for 28 days, on which they sustained maximum growth. Group II (protein-depleted) was fed for 14 days on 0.5 lactalbumin diet, which caused loss of weight; this was followed by repletion for 14 days with the control diet. Group III (protein-energy restricted) received a 1% lactalbumin diet at one-half the food intake of group II for 14 days, and this was also followed by 14 days of repletion with the control diet. 3. The controls showed a progressive rise in the daily urinary output of N-tau-methylhistidine, which was proportionally slightly less rapid than the body-weight increase. 4. The protein-depleted group II showed a marked and progressive decrease in N-tau-methylhistidine excretion, which was proportionally greater than the fall in body weight; during repletion, N-tau-methylhistidine output rose in parallel with body-weight increase, but it did not reach the value attained by the control group. 5. Group III, restricted in both dietary protein and energy, showed an initial small increase in daily N-tau-methylhistidine output, which contrasted with the sharp loss of body weight during this period. After 11 days on this restricted diet, group III then underwent a decrease in N-tau-methylhistidine output, which persisted into the first 4 days of the repletion period, after which output of the methylated amino acid became the same as for group II. 6. Creatinine output, used as an additional metabolic measure of muscle metabolism, showed a fairly constant relationship to body weight in groups I and II during depletion and repletion. However, rats with protein-energy deficiency (group III) underwent a marked increase in output of creatinine per unit of body weight, which also persisited into the repletion period before it fell to more normal values relative to body weight. 7. Analysis of the N-tau-methylhistidine content of actin isolated from a group of protein-depleted rats revealed a small (5%) but significance (P less than 0.02) decrease relative to well-nourished controls. 8. Hence, the rate of muscle protein degradation, as indicated by changes in urinary N-tau-methylhistidine output, appears to respond sensitively and in opposite directions to insufficiency of protein of energy in the diet.

Ntau-methylhistidine content of mixed proteins in various rat tissues.

In order to use Ntau-methylhistidine (3-methylhistidine) excretion in the urine as a measure of muscle protein breakdown, it is necessary to demonstrate that other tissues are not important sources of this protein constituent. Accordingly, the concentration of Ntau-methylhistidine in blood serum and in the mixed proteins of heart, brain, lung, kidney, diaphragm, spleen, testis, stomach, liver and hind leg skeletal muscle was measured in male rats of approx. 400 g body weight. The free Ntau-methylhistidine concentration of rat serum was less than 2 nmol per ml. In contrast, measurable amounts of Ntau-methylhistidine were found in the mixed proteins of all tissues and organs examined. The highest concentration was found in skeletal muscle (658 nmol/g tissue). Assuming muscle mass to be 45% of body weight, it has been estimated that the muscle contains more than ten times the total amount of this amino acid present in all of the other organs analyzed, which together account for about 20% of total body weight. These findings indicate that skeletal muscle is likely to be the major source of urinary Ntau-methylhistidine and the latter is, in consequence, a reflection of myofibrillar protein breakdown in skeletal muscle.

Metabolism of 3-methylhistidine in man.

The metabolism of L-3-methylhistidine was studied in man using intravenously administered ((14)C)3-methylhistidine. Analysis for expired (14)CO2 for periods up to 2 hr following a single intravenous injection revealed no radioactivity, indicating that this compound is not oxidized in man. Analysis of urine samples for total radioactivity showed that 75% of the administered dose was excreted in 24 hr and 95% in 48 hr. Ion-exchange chromatography of urine samples with monitoring of the column eluated by a flow liquid-scintillation technique showed the presence of only two radioactive peaks. The time taken to elute these peaks was compatible with the major excretory component (95.5%) being ((14)C)3-methylhistidine, accompanied by a small amount (4.5%) in the form of N-acetyl-((14)C)3-methylhistidine. The plasma disappearance curves of ((14)C)3-methylhistidine suggested a half-life of approximately 130 min. The inability ot oxidize 3-methylhistidine and its quantitative excretion as the original compound as well as its N-acetyl derivative is similar to its metabolic fate in the rat and therefore suggests that 3-methylhistidine excretion may provide a reliable measure of actin and myosin turnover in the whole animal or in human subjects.