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. 1980 Jun 15;188(3):705–713. doi: 10.1042/bj1880705

Enzymic determination of branched-chain amino acids and 2-oxoacids in rat tissues. Transfer of 2-oxoacids from skeletal muscle to liver in vivo.

G Livesey, P Lund
PMCID: PMC1161952  PMID: 6781474

Abstract

1. A procedure is described for the purification of leucine dehydrogenase (EC 1.4.1.9) from Bacillus subtilis. 2. The preparation is suitable for the quantitative assay of branched-chain amino acids and their 2-oxoacid analogues. 3. The content of total branched-chain 2-oxoacids in freeze-clamped liver, kidney, heart or mammary gland of fed rats is less than 5 nmol/g fresh wt. Higher amounts are present in skeletal muscle and arterial blood (25 +/- 4 nmol per g fresh wt., and 33 +/- 6 nmol per ml respectively; means +/- S.D. of 3 and 11 animals respectively). The values are not significantly affected by starvation for 24 h. 4. Arteriovenous difference measurements show that considerable amounts of branched-chain 2-oxoacids are released by skeletal muscle into the circulation and similar amounts are removed by the liver (about 1 mmol/24 h in a 400 g rat).

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chang T. W., Goldberg A. L. Leucine inhibits oxidation of glucose and pyruvate in skeletal muscles during fasting. J Biol Chem. 1978 May 25;253(10):3696–3701. [PubMed] [Google Scholar]
  2. Chang T. W., Goldberg A. L. The metabolic fates of amino acids and the formation of glutamine in skeletal muscle. J Biol Chem. 1978 May 25;253(10):3685–3693. [PubMed] [Google Scholar]
  3. Chatwin A. L., Linzell J. L., Setchell B. P. Cardiovascular changes during lactation in the rat. J Endocrinol. 1969 Jun;44(2):247–254. doi: 10.1677/joe.0.0440247. [DOI] [PubMed] [Google Scholar]
  4. Cree T. C., Hutson S. M., Harper A. E. Gas-liquid chromatography of alpha-keto acids: quantification of the branched-chain-alpha-keto acids from physiological sources. Anal Biochem. 1979 Jan 1;92(1):159–163. doi: 10.1016/0003-2697(79)90638-9. [DOI] [PubMed] [Google Scholar]
  5. Danner D. J., Lemmon S. K., Elsas L. J., 2nd Substrate specificity and stabilization by thiamine pyrophosphate of rat liver branched chain alpha-ketoacid dehydrogenase. Biochem Med. 1978 Feb;19(1):27–38. doi: 10.1016/0006-2944(78)90004-2. [DOI] [PubMed] [Google Scholar]
  6. Davis E. J., Bremer J. Studies with isolated surviving rat hearts. Interdependence of free amino acids and citric-acid-cycle intermediates. Eur J Biochem. 1973 Sep 21;38(1):86–97. doi: 10.1111/j.1432-1033.1973.tb03037.x. [DOI] [PubMed] [Google Scholar]
  7. Garber A. J., Karl I. E., Kipnis D. M. Alanine and glutamine synthesis and release from skeletal muscle. II. The precursor role of amino acids in alanine and glutamine synthesis. J Biol Chem. 1976 Feb 10;251(3):836–843. [PubMed] [Google Scholar]
  8. Goldstein L., Newsholme E. A. The formation of alanine from amino acids in diaphragm muscle of the rat. Biochem J. 1976 Feb 15;154(2):555–558. doi: 10.1042/bj1540555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hermier J., Lebeault J. M., Zévaco C. Purification et propriétés des déshydrogénases a NAD intervenant dans la germination de la spore de Bacillus subtilis. II. L. leucine: NAD oxydo-réductase (désaminate) Bull Soc Chim Biol (Paris) 1970;52(10):1089–1104. [PubMed] [Google Scholar]
  10. Hutson S. M., Cree T. C., Harper A. E. Regulation of leucine and alpha-ketoisocaproate metabolism in skeletal muscle. J Biol Chem. 1978 Nov 25;253(22):8126–8133. [PubMed] [Google Scholar]
  11. Krebs H. A., Lund P. Aspects of the regulation of the metabolism of branched-chain amino acids. Adv Enzyme Regul. 1976;15:375–394. doi: 10.1016/0065-2571(77)90026-7. [DOI] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Lund P., Baverel G. Interference by branched-chain amino acids in the assay of alanine with alanine dehydrogenase. Biochem J. 1978 Sep 15;174(3):1079–1082. doi: 10.1042/bj1741079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Odessey R., Goldberg A. L. Leucine degradation in cell-free extracts of skeletal muscle. Biochem J. 1979 Feb 15;178(2):475–489. doi: 10.1042/bj1780475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ohshima T., Misono H., Soda K. Properties of crystalline leucine dehydrogenase from Bacillus sphaericus. J Biol Chem. 1978 Aug 25;253(16):5719–5725. [PubMed] [Google Scholar]
  16. Parker P. J., Randle P. J. Partial purification and properties of branched-chain 2-oxo acid dehydrogenase of ox liver. Biochem J. 1978 Jun 1;171(3):751–757. doi: 10.1042/bj1710751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pettit F. H., Yeaman S. J., Reed L. J. Purification and characterization of branched chain alpha-keto acid dehydrogenase complex of bovine kidney. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4881–4885. doi: 10.1073/pnas.75.10.4881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Snell K., Duff D. A. The release of alanine by rat diaphragm muscle in vitro. Biochem J. 1977 Feb 15;162(2):399–403. doi: 10.1042/bj1620399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Soda K., Misono H., Mori K., Sakato H. Crystalline L-leucine dehydrogenase. Biochem Biophys Res Commun. 1971 Aug 20;44(4):931–935. doi: 10.1016/0006-291x(71)90801-1. [DOI] [PubMed] [Google Scholar]
  20. Spydevold O. Sources of carbon skeleton of alanine released from skeletal muscle. Acta Physiol Scand. 1976 Jul;97(3):273–280. doi: 10.1111/j.1748-1716.1976.tb10264.x. [DOI] [PubMed] [Google Scholar]
  21. WILLIAMSON D. H., MELLANBY J., KREBS H. A. Enzymic determination of D(-)-beta-hydroxybutyric acid and acetoacetic acid in blood. Biochem J. 1962 Jan;82:90–96. doi: 10.1042/bj0820090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. WOLLENBERGER A., RISTAU O., SCHOFFA G. [A simple technic for extremely rapid freezing of large pieces of tissue]. Pflugers Arch Gesamte Physiol Menschen Tiere. 1960;270:399–412. [PubMed] [Google Scholar]
  23. Walser M., Lund P., Ruderman N. B., Coulter A. W. Synthesis of essential amino acids from their alpha-keto analogues by perfused rat liver and muscle. J Clin Invest. 1973 Nov;52(11):2865–2877. doi: 10.1172/JCI107483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wohlhueter R. M., Harper A. E. Coinduction of rat liver branched chain alpha-keto acid dehydrogenase activities. J Biol Chem. 1970 May 10;245(9):2391–2401. [PubMed] [Google Scholar]
  25. YOSHIDA A., FREESE E. PURIFICATION AND CHEMICAL CHARACTERIZATION OF ALANINE DEHYDROGENASE OF BACILLUS SUBTILIS. Biochim Biophys Acta. 1964 Oct 23;92:33–43. doi: 10.1016/0926-6569(64)90266-4. [DOI] [PubMed] [Google Scholar]

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