Measurement of protein turnover in normal man using the end-product method with oral [15N]glycine: comparison of single-dose and intermittent-dose regimens

Abstract

The ‘single-dose end-product’ approach for the measurement of protein turnover is the only method suited for application to free-living individuals and for field studies. However, the method has never been widely accepted because the results obtained appear to vary from one situation to another. There is the need for a formal comparison of the different approaches that have been used and the assumptions on which they are based in order to be able to understand the extent to which the variation in the results obtained is real or might be attributed to methodological differences. The present study used the ‘prime/intermittent-dose end-product’ approach over 18 h as a reference with which to compare the ‘single-dose end-product’ approach, with urine being collected for periods of 9 to 48 h. N flux was derived in a total of thirteen normal men using oral [15N]glycine and measurement ofI5N enrichment in urinary NH3or urea, with isotope being given either as prime/intermittent doses or a single dose in separate studies. The pattern of results was similar to those reported in the literature. In all studies the rates of N flux derived from urea were higher than those derived from NH3, under equivalent conditions, by about 25–30%. The single-dose approach, with urine collection over 9 h, gave results which were consistently higher than the prime/intermittent-dose approach. The extent of the difference was influenced by the duration of time over which the cumulative excretion of isotope in urine was determined, and with NH3as the end-product the most consistent estimates of N flux could be obtained with a urine collection of at least 12 h and no greater than 24 h. With urea as the end-product, correction for the label retained in the body pool at 9 h gave similar results for N flux to those derived from the total excretion of label in urea over 24 h. The derivation of values for protein synthesis and protein degradation from measures of flux requires care to ensure that the time periods over which N intake and excretion are measured accord with those for which the measurement of flux apply. It is concluded that measurements of protein turnover similar to those obtained with the prime/intermittent-dose approach can be obtained with the single-dose approach in the fed state during the daytime, either from the excretion of label in urinary NH3over a period of 12 h or with the excretion of label in urinary urea over a period of 24 h. The suitability of the method for use in the fasted state or at night time remains to be determined.