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Amino Acids

, Volume 7, Issue 3, pp 267–278 | Cite as

Source of amino acids for tRNA acylation in growing chicks

  • D. M. Barnes
  • C. C. Calvert
  • K. C. Klasing
Article

Summary

Specific radioactivity in three amino acid compartments was examined in broiler chicks following a flooding dose of leucine or phenylalanine. In general, specific radioactivity of leucine and phenylalanine in deproteinated plasma (SAe) and tissue (SAi) compartments, exceeded that in acylated-tRNA (SAt). In most tissues, SAe and SAi rapidly reached a similar peak level by 5 min followed by a slow decline for the next 30 minutes. Many tissues (eg. GI tract, liver, skin, and thigh) failed to maintain equilibrium between SAe and SAi over time. More metabolically active tissues, such as GI and liver had the greatest differences between these compartments. The difference between SAe and SAi for both leucine and phenylalanine were due to SAi decreasing faster than SAe, indicating dilution with unlabelled amino acids from proteolysis. Plasma and tissue specific radioactivity overestimated tRNA specific radioactivity by as much as 5 and 2.8 fold using leucine or 2.7 and 1.4 fold using phenylalanine, respectively. These data suggest that intracellular compartmentation of protein metabolism and the coupling of protein degradation and synthesis occur, in vivo.

Keywords

Amino acids Protein synthesis tRNA charging Amino acid metabolism 

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References

  1. Airhart J, Vidrich A, Khairallah EA (1974) Compartmentation of free amino acids for protein synthesis. Biochem J 140: 539–545Google Scholar
  2. Airhart J, Arnold JA, Bulman CA, Low RB (1981) Protein synthesis in pulmonary alveolar macrophages. Biochim Biophys Acta 653: 108–117Google Scholar
  3. Barnes DM, Calvert CC, Klasing KC (1992) Source of amino acids for tRNA acylation: implications for measurement of protein synthesis. Biochem J 283: 583–589Google Scholar
  4. Bernier JF, Calvert CC, Famula TR, Baldwin RL (1986) Maintenance energy requirement and net energetic efficiency in mice with a major gene for rapid postweaning gain. J Nutr 116: 419–428Google Scholar
  5. Bidlingmeyer BA, Cohen SA, Tarvin TL (1984) Rapid analysis of amino acids using precolumn derivatization. J Chromatogr 336: 93–104Google Scholar
  6. Chikenji MD, Elwyn DH, Kinney JM (1983) Protein synthesis rates in rat muscle and skin based on lysyl-tRNA radioactivity. J Surg Res 34: 68–82Google Scholar
  7. Garlick PJ, McNurlan MA, Preedy VR (1980) A rapid and convenient technique for measuring the rate of protein synthesis in tissues by injection of [3H]phenylalanine. Biochem J 192: 719–723Google Scholar
  8. Hall GE, Yee JA (1989) Parathyroid hormone alteration of free and tRNA-bound proline specific activities in cultured mouse osteoblast-like cells. Biochem Biophys Res Comm 161: 994–1000Google Scholar
  9. Hammer JA, Rannels DE (1981) Protein turnover in pulmonary macrophages. J Biol Chem 246: 435–446Google Scholar
  10. Henshaw EC, Hirche CA, Morton BE, Hiatt HH (1971) Control of protein synthesis in mammalian tissues through changes in ribosome activity. J Biol Chem 246: 435–446Google Scholar
  11. Hider RC, Fern EB, London DR (1971) Identification in skeletal muscle of a distinct extracellular pool of amino acids, and its role in protein synthesis. Biochem J 121: 817–827Google Scholar
  12. Kelley J, Stirewalt WS, Chrin L (1984) Protein synthesis in rat lung: measurements in vivo based on leucyl-tRNA and rapidly turning-over procollagen I. Biochem J 222: 77–83Google Scholar
  13. Khairallah EA, Mortimore GE (1976) Assessment of protein turnover in perfused rat liver. Evidence for amino acid compartmentation from differential labeling of free and tRNA-bound valine. J Biol Chem 251: 1375–1384Google Scholar
  14. Klasing KC, Austic RE (1984) Changes in plasma, tissue and urinary nitrogen metabolites due to an inflammatory challenge. Proc Soc Exp Biol Med 176: 276–284Google Scholar
  15. McKee EE, Cheung JY, Rannels DE, Morgan HE (1978) Measurement of the rate of protein synthesis and compartmentation of heart phenylalanine. J Biol Chem 253: 1030–1040Google Scholar
  16. McNurlan MA, Tomkins AM, Garlick PJ (1979) The effect of starvation on the rate of protein synthesis in rat liver and small intestine. Biochem J 178: 373–379Google Scholar
  17. Mirande M, Le Corre D, Louvard D, Reggio H, Pailliez JP, Waller JP (1985) Association of an aminoacyl-tRNA synthetase with the cytoskeletal framework fraction from mammalian cells. Exp Cell Res 156: 91–102Google Scholar
  18. Moline G, Hampel A, Enger MD (1974) Polyribosomal and particulate distribution of lysyland phenylalanyl-transfer ribonucleic acid synthetase. Biochem J 143: 191–195Google Scholar
  19. Negrutskii BS, Deutscher MP (1991) Channeling of aminoacyl-tRNA for protein synthesis in vivo. Proc Natl Acad Sci USA 88: 4991–4995Google Scholar
  20. Obled C, Barre F, Millward DJ, Arnal M (1989) Whole body protein synthesis: studies with different amino acids in the rat. Am J Physiol 257: E639-E646Google Scholar
  21. Opsahl WP, Ehrhart LA (1987) Compartmentalization of proline pools and apparent rates of collagen and non-collagen protein synthesis in arterial smooth muscle cells in culture. Biochem J 243: 137–144Google Scholar
  22. Schaefer AL, Scott SL (1993) Amino acid flooding doses for measuring rates of protein synthesis. Amino Acids 4: 5–20Google Scholar
  23. Schneible PA, Airhart J, Low RB (1981) Differential compartmentation of leucine for oxidation and for protein synthesis in cultured skeletal muscle. J Biol Chem 256: 4888–4894Google Scholar
  24. Schneible PA, Young RB (1984) Leucine pools in normal and dystrophic chicken skeletal muscle cells in culture. J Biol Chem 259: 1436–1440Google Scholar
  25. Seglen PO, Gordon PB, Poli A (1980) Amino acid inhibition of the autophagic/lysosomal pathway of protein degradation in isolated rat hepatocytes. Biochem Biophys Acta 630: 103–118Google Scholar
  26. Sivaram P, Deutscher MP (1990) Existance of two forms of rat arginyl-tRNA synthestase suggests chnneling of aminoacyl-tRNA for protein synthesis. Proc Natl Acad Sci USA 87: 3665–3669Google Scholar
  27. Smith CB, Sun Y, Deibler GE, Sokoloff L (1991) Effect of loading doses of L-valine on relative contributions of valine derived from protein degradation and plasma to the precursor pool for protein synthesis in rat brain. J Neurochem 57: 1540–1547Google Scholar
  28. Southorn BG, Kelly JM, McBride BW (1992) Phenylalanine flooding dose procedure is effective in measuring intestinal and liver protein synthesis in sheep. J Nutr 122: 2398–2407Google Scholar
  29. Stirewalt WS, Low RB (1983) Effects of insulin in vivo on protein turnover in rat epitrochlearis muscle. Biochem J 210: 323–330Google Scholar
  30. Sun Y, Deibler GE, Sokoloff L, Smith CB (1992) Determination of regional rates of cerebral protein synthesis adjusted for regional differences in recycling of leucine derived from protein degradation into the precursor pool in conscious adult rats. J Neurochem 59: 863–873Google Scholar
  31. Tischler ME, Desautels M, Goldberg AL (1982) Does leucine, leucyl-tRNA or some metabolite of leucine regulate protein synthesis and degradation in skeletal and cardiac muscle? J Biol Chem 257: 1613–1624Google Scholar
  32. Watt PW, Lindsay Y, Scrimgeour CM, Chien PAF, Gibson JNL, Taylor DJ, Rennie MJ (1991) Isolation of aminoacyl-tRNA and its labeling with stable-isotope tracers: use in studies of human tissue protein synthesis. Proc Natl Acad Sci 88: 5892–5896Google Scholar
  33. Zak R, Martin AF, Blough R (1979) Assessment of protein turnover by use of radioisotopic tracers. Physiol Rev 59: 407–447Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • D. M. Barnes
    • 1
  • C. C. Calvert
    • 2
  • K. C. Klasing
    • 1
  1. 1.Department of Avian ScienceUniversity of CaliforniaDavisUSA
  2. 2.Department of Animal ScienceUniversity of CaliforniaDavisUSA

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