The measurement of tissue protein turnover.
Tissue protein turnover can be assessed by a number of semi-, quantitative and qualitative methods. There are a number of static indices of the state of turnover of protein, for example amount of RNA per DNA or protein, the state of aggregation of ribosomes (i.e. the polyribosome index), the abundance of mRNA for particular proteins, and the enzymatic activity of proteins such as proteases, ribonuclease, etc. In addition, the concentration of particular amino acids such as glutamine or non-re-utilizable amino acids, formed post-translationally, such as 3-methylhistidine or hydroxyproline, are able to provide snapshot indices. However, since turnover is a dynamic process it should, ideally, be probed using methods such as the incorporation of tracer amino acids into protein or the dilution of tracer amino acids in the free pool by protein breakdown. The combination of tracer and tissue or limb balance methods is especially powerful since all the dynamic processes can potentially be quantified. The use of stable isotopes to label metabolic tracers has dramatically increased the feasibility of carrying out measurements of protein synthesis and breakdown and there has been a substantial growth in the application of the methods to a wide variety of tissues sampled by biopsy or at operation. Summaries of a number of currently feasible methods are provided, together with commentary on the relative efficacy of the methods and of the instrumental techniques required. There is also a discussion of suitable tracer labels and amino acids, plus a summary of the most reliable current values for protein turnover in a variety of tissues. The review also contains descriptions of potential methods which have not yet been applied in human beings but which are feasible, given the current recent increases in the accuracy and sensitivity of instrumentation for measurement of stable isotope labelling.
Smith
K
K
Department of Anatomy & Physiology, University of Dundee, UK.
Rennie
M J
MJ
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D006801:Humans; D007700:Kinetics; D009696:Nucleic Acids / Q000378:metabolism; D011506:Proteins / Q000032:analysis / Q000378:metabolism*
D000596:Amino Acids; D009696:Nucleic Acids; D011506:Proteins