Hostname: page-component-cd9895bd7-7cvxr Total loading time: 0 Render date: 2024-12-24T01:36:56.241Z Has data issue: false hasContentIssue false
  • English
  • Français

Valine oxidation: the synthesis and evaluation of l-[3-3H]valine as a tracer in vivo

Published online by Cambridge University Press:  09 March 2007

P. R. Beckett ,
A. Cadenhead  and
M. F. Fuller
P. R. Beckett
Affiliation:
Rowett Research Institute, Bucksburn, Aberdeen AB2 9SB
A. Cadenhead
Affiliation:
Rowett Research Institute, Bucksburn, Aberdeen AB2 9SB
M. F. Fuller
Affiliation:
Rowett Research Institute, Bucksburn, Aberdeen AB2 9SB
Rights & Permissions [Opens in a new window]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

The suitability of l-[3-3H]valine for measuring valine oxidation was studied by comparing its oxidation rate with that of l-[1-14C]valine in rats and pigs. l-[3-3H]valine was synthesized by removal of the tritium on carbon-2 of l-[2,3-3H]valine by acetylation. The acetyl group was removed enzymatically using pig renal acylase 1 (EC 3.5.1.14) and the product was purified by ion-exchange and paper chromatography. For the first rat experiment l-[3-3H]valine was synthesized in our laboratory; for the subsequent experiments it was produced by Amersham International plc. In the first experiment in rats the two tracers were given by injection and 14CO2 was collected for 2 h. The oxidation of tritiated valine was significantly higher than that of l-[1-14C]valine. In a second experiment there was no difference. This was probably due to the higher purity of the labelled valine which, for the second experiment, was shown by nuclear magnetic resonance to contain only one tritium atom. In a study with pigs in which the two tracers were given by continuous infusion there was no significant difference between them in flux or oxidation. The results of this experiment were used to evaluate a model to estimate amino acid requirements. With pigs given a methionine-limiting diet a reduction in methionine intake, by reducing protein accretion, increased valine oxidation by the same proportion.

Keywords

Tritiated valineMethionine/valine requirementPig
Type
Amino Acid Metabolism
Information
British Journal of Nutrition , Volume 68 , Issue 1 , July 1992 , pp. 139 - 151
Copyright
Copyright © The Nutrition Society 1992

References

REFERENCES

Adriaens, P., Meesschart, B. & Vanderhaeghe, H. (1975). Isotope distribution in l-valine-2,3-T. Analytical Biochemistry 69, 297–299.CrossRefGoogle ScholarPubMed
Ball, R. O. & Bayley, H. S. (1986). Influence of dietary protein concentration on the oxidation of phenylalanine by the young pig. British Journal of Nutrition 55, 651–658.CrossRefGoogle ScholarPubMed
Beckett, P. R. (1989). Amino acid oxidation. PhD Thesis, University of Aberdeen.Google Scholar
Bergner, H. & Simon, O. (1976). Methodische Untersuchungen zur stoffwechselorientieren Aminosäuren-bedarfsbestimmung bei Ratten im Erhaltungszustand anhand der Oxydationsrate von 14C-markiertem Lysin zu 14CO2 (Studies investigating methods for determining the amino acid requirements on the basis of metabolic data through measurements of oxidation of CO2). Archiv für Tierernährung 26, 815–826.CrossRefGoogle Scholar
Bergner, H., Simon, O. & Adam, K. (1978). Lysinbedarfsbestimmung bei wachsenden Ratten anhand der Katabolisierungsrate von 14C- and 15N-markiertem Lysin (Lysine requirement determination on growing rats by means of the catabolization rates of 14C- and 15N-labelled lysine). Archiv für Tierernährung 28, 21–29.CrossRefGoogle Scholar
Brookes, I. M., Owens, F. N. & Garrigus, U. S. (1972). Influence of amino acid level in the diet upon amino acid oxidation by the rat. Journal of Nutrition 102, 27–36.CrossRefGoogle ScholarPubMed
Crawhall, J. C. & Smyth, D. G. (1958). The synthesis of tritiovaline and its incorporation into rat-visceral proteins. Biochemical Journal 69, 280–286.CrossRefGoogle ScholarPubMed
Culebras, J. M., Fitzpatrick, G. F., Brennan, M. F., Boyden, C. M. & Moore, F. D. (1977). Total body water and the exchangeable hydrogen. 11. A review of comparative data from animals based on isotope dilution and desiccation with a report of new data from the rat. American Journal of Physiology 232, R60–R65.Google Scholar
Fuller, M. F., McWilliam, R., Wang, T. C. & Giles, L. R. (1989). The optimum dietary amino acid pattern for growing pigs. 2. Requirements for maintenance and for tissue protein accretion. British Journal of Nutrition 62, 255–267.CrossRefGoogle ScholarPubMed
Fuller, M. F., Reeds, P. J., Cadenhead, A., Seve, B. & Preston, T. (1987). Effects of the amount and quality of dietary protein on nitrogen metabolism and protein turnover in pigs. British Journal of Nutrition 58, 287–300.CrossRefGoogle Scholar
Greenstein, J. P. & Winitz, M. (1961). Chemistry of the Amino Acids. London: Wiley.Google Scholar
Haggarty, P., Reeds, P. J., Fletcher, J. M. & Wahle, K. W. J. (1986). The fate of 14C derived from radioactively labelled dietary precursors in young rats of the Zucker strain (Fa/- and fa/fa). Biochemical Journal 235, 323–327.CrossRefGoogle ScholarPubMed
Harper, A. E., Miller, R. H. & Block, K. P. (1984). Branched-chain amino acid metabolism. Annual Review of Nutrition 4, 409–545.CrossRefGoogle ScholarPubMed
Kang-Lee, Y. A. & Harper, A. E. (1978). Threonine metabolism in vivo: effect of threonine intake and prior induction of threonine dehydratase in rats. Journal of Nutrition 108, 163–171.CrossRefGoogle ScholarPubMed
Kim, K., Elliot, J. I. & Bayley, H. S. (1983). Oxidation of an indicator amino acid by young pigs receiving diets with varying levels of lysine or threonine, and an assessment of amino acid requirements. British Journal of Nutrition 50, 391–401.CrossRefGoogle ScholarPubMed
Lobley, G. E., Connell, A. & Watson, B. T. (1985). The use of a proprietary organic sample oxidiser to determine 14CO2 and 14C-carbonates of trace level specific radioactivity. International Journal of Applied Radiation and Isotopes 36, 905–907.CrossRefGoogle ScholarPubMed
Mitchell, A. R., Kent, S. B. H., Chu, I. C. & Merrifield, R. B. (1978). Quantitative determination of d- and l-amino acids by reaction with tert-butyloxycarbonyl-l-leucine N-hydroxysuccinimide ester and chromatographic separation as l,d and l,l dipeptides. Analytical Chemistry 50, 637–640.CrossRefGoogle Scholar
Mitchell, H. H. (1950). Some species differences in amino acid requirements. In Protein and Amino Acid Requirements of Mammals [Albanese, A. A., editor]. New York: Academic Press.Google Scholar
Mitz, M. A. & Schlueter, R. J. (1958). Direct spectrophotometric measurement of the peptide bond: application to the determination of acylase I. Biochimica et Biophysica Acta 27, 168–172.CrossRefGoogle Scholar
Newport, M. J., Chavez, E. R., Horney, F. D. & Bayley, H. S. (1976). Amino acid metabolism in the piglet: influence of level of protein and of methionine in the diet on tissue uptake and in vivo oxidation. British Journal of Nutrition 36, 87–99.CrossRefGoogle ScholarPubMed
Pichat, L., Rochas, G. & Herbert, M. (1965). Préparation de l'acide dl-α aminoadipique 14C-6 à partir de cyanure de potassium 14C et à benzamido δ-valero-acetone (Synthesis of dl-α-aminoadipic acid-6-14C from KCN-14C and α-benzamido-δ-valerolactone). Bulletin. Société Chimique de France 5, 1384–1386.Google Scholar
Reeds, P. J., Cadenhead, A., Fuller, M. F., Lobley, G. E. & McDonald, J. D. (1980). Protein turnover in growing pigs. Effects of age and food intake. British Journal of Nutrition 43, 445–455.CrossRefGoogle ScholarPubMed
Reeds, P. J., Fuller, M. F., Cadenhead, A., Lobley, G. E. & McDonald, J. D. (1981). Effects of changes in the intakes of protein and non-protein energy on whole-body protein turnover in growing pigs. British Journal of Nutrition 45, 539–546.CrossRefGoogle ScholarPubMed
Rosenhagen, M. & Segal, S. (1974). Stereospecificity of amino acid uptake by rat and human kidney cortex slices. American Journal of Physiology 227, 843–847.CrossRefGoogle ScholarPubMed
Soliman, A.-G. & Harper, A. E. (1971). Effect of protein content of diet of lysine oxidation by the rat. Biochimica et Biophysica Acta 244, 146–154.CrossRefGoogle ScholarPubMed
Staten, M. A., Bier, D. M. & Mathews, D. E. (1984). Regulation of valine metabolism in man: a stable isotope study. American Journal of Clinical Nutrition 40, 1224–1234.CrossRefGoogle Scholar
Tanaka, K., Aberhart, D. J., Amster, J. I., Jenkins, E. E. & Hsu, C.-T. (1981). Metabolism of valine chirally labelled with carbon 13 and deuterium in vitamin b12-folate-deficient rate in vivo. Studies on the physiological role of keto-enol tautomerization. Journal of Biological Chemistry 255, 7763–7768.CrossRefGoogle Scholar
Tomera, J. F., McCaffrey, P. G. & Brunengraber, H. (1983). Rates of catabolism calculated from 14CO2 production: artifacts and realities. Pharmacology, Biochemistry and Behaviour 18, 285–288.CrossRefGoogle ScholarPubMed
Wang, T. C. & Fuller, M. F. (1989). The optimum dietary amino acid pattern for growing pigs. 1. Experiments by amino acid deletion. British Journal of Nutrition 62, 77–89.CrossRefGoogle ScholarPubMed
Waterlow, J. C., Garlick, P. J. & Millward, D. J. (1978). Protein Turnover in Mammalian Tissues and in the Whole Body. Oxford: North-Holland Publishing Co.Google Scholar