Use of the constant infusion technique for measuring rates of protein synthesis in the New Zealand White rabbit

Abstract

1.To study the potential of the constant-infusion technique for measuring rates of protein synthesis in New Zealand White rabbits, animals were infused for up to 6 h with radioactively-labelled tyrosine.2.Labelled tyrosine from plasma and tissues was isolated from labelled metabolites by ion-exchange chromatography.3.Analysis of serial blood and muscle biopsy samples removed under anaesthesia showed that the specific radioactivity (SR) of the free tyrosine pools reached an approximately constant value within 2 h.4.Certain commercial preparations of L-[side-chain 2,3-3H]tyrosine were contaminated with 300 mg radioactive D-tyrosine/g. The D-isomer appeared to enter the muscle intracellular pool.5.In constant-infusion experiments L-[3H]tyrosine could replace the uniformly-14C-labelled L-isomer for the determination of rates of protein synthesis in muscle. L-[side-chain 2,3-3H]tyrosine may not be suitable for use as a precursor for measuring rates of liver protein synthesis.6.Evidence is presented that the precursor of liver protein synthesis may not be well defined by the SR for free tyrosine of the homogenate.7.The technique was used to measure the rates of protein synthesis in adult rabbits. The rates of protein synthesis in liver and muscle were measured and from measurements of tyrosine flux the mean rate of whole-body protein synthesis was calculated as 13.8 g/kg per d.