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Am J Physiol Endocrinol Metab 282: E1029-E1038, 2002. First published January 22, 2002; doi:10.1152/ajpendo.00215.2001
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Vol. 282, Issue 5, E1029-E1038, May 2002

Aminoacyl-tRNA enrichment after a flood of labeled phenylalanine: insulin effect on muscle protein synthesis

Giuseppe Caso1, G. Charles Ford2, K. Sreekumaran Nair2, Peter J. Garlick1, and Margaret A. McNurlan1

1 Department of Surgery, State University of New York, Stony Brook, New York 11794-8191; and 2 Endocrine Research Unit, Division of Endocrinology and Metabolism, Mayo Clinic and Mayo Foundation, Rochester, Minnesota 55905

Muscle protein synthesis in dogs measured by flooding with L-[2H5]phenylalanine (70 mg/kg) was significantly stimulated by infusion of insulin with amino acids. The stimulation of muscle protein synthesis was similar when calculated from the enrichment of phenylalanyl-tRNA (61 ± 10%, P < 0.001), plasma phenylalanine (61 ± 10%, P < 0.001), or tissue fluid phenylalanine (54 ± 10%, P < 0.001). The time course for changes in enrichment of L-[2H5]phenylalanine throughout the flooding period was determined for plasma, tissue fluid, and phenylalanyl-tRNA in the basal state and during the infusion of insulin with amino acids. Enrichments of plasma free phenylalanine and phenylalanyl-tRNA were equalized between 20 and 45 min, although the enrichment of phenylalanyl-tRNA was lower at early time points. Rates of muscle protein synthesis obtained with the flooding method and calculated from plasma phenylalanine enrichment were comparable to those calculated from phenylalanyl-tRNA and also to those obtained previously with a continuous infusion of phenylalanine with phenylalanyl-tRNA as precursor. This study confirms that, with a bolus injection of labeled phenylalanine, the enrichment of aminoacyl-tRNA, the true precursor pool for protein synthesis, can be assessed from more readily sampled plasma phenylalanine.

amino acids; L-[2H5]phenylalanine; dog; flooding technique; ketoisocaproic acid; stable isotopes


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