Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1975 Nov;152(2):401–408. doi: 10.1042/bj1520401

The activity of phosphoenolpyruvate carboxykinase in rat tissues. Assay techniques and effects of dietary and hormonal changes

Christopher I Pogson 1, Stephen A Smith 1
PMCID: PMC1172484  PMID: 1220693

Abstract

1. Phosphoenolpyruvate carboxykinase was assayed by three methods: (i) incorporation of H14CO3− into oxaloacetate: (ii) conversion of oxaloacetate into phosphoenolpyruvate, subsequently assayed enzymically; and (iii) transfer of 32P from [γ-32P]GTP to oxaloacetate. 2. Enzyme activity is increased in liver and epididymal adipose tissue in alloxan-diabetes and starvation, and in kidney in starved, acidotic and steroid-treated animals. 3. The ratios of the `back' to the `forward' reactions in liver, kidney and epididymal adipose tissue are different and characteristic of each tissue; they differ markedly from values reported for the purified mitochondrial enzyme. 4. The ratio of the `back' to `forward' reaction in any one tissue is constant in adrenalectomized, diabetic, acidotic and steroid-treated animals. 5. In starved animals, the ratio is increased in liver and kidney, but decreased in epididymal adipose tissue. 6. Administration of l-tryptophan results in an acute (1h) increase in activity measured in the `forward' direction alone in liver and epididymal adipose tissue, but not in kidney.

Full text

PDF
401

Selected References

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

  1. Alleyne G. A., Scullard G. H. Renal metabolic response to acid base changes. I. Enzymatic control of ammoniagenesis in the rat. J Clin Invest. 1969 Feb;48(2):364–370. doi: 10.1172/JCI105993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson J. W., Stowring L. Glycolytic and gluconeogenic enzyme activities in renal cortex of diabetic rats. Am J Physiol. 1973 Apr;224(4):930–936. doi: 10.1152/ajplegacy.1973.224.4.930. [DOI] [PubMed] [Google Scholar]
  3. Ballard F. J., Hanson R. W. Phosphoenolpyruvate carboxykinase and pyruvate carboxylase in developing rat liver. Biochem J. 1967 Sep;104(3):866–871. doi: 10.1042/bj1040866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ballard F. J., Hanson R. W. Purification of phosphoenolpyruvate carboxykinase from the cytosol fraction of rat liver and the immunochemical demonstration of differences between this enzyme and the mitochondrial phosphoenolpyruvate carboxykinase. J Biol Chem. 1969 Oct 25;244(20):5625–5630. [PubMed] [Google Scholar]
  5. Ballard F. J., Hopgood M. F. Phosphopyruvate carboxylase induction by L-tryptophan. Effects on synthesis and degradation of the enzyme. Biochem J. 1973 Oct;136(2):259–264. doi: 10.1042/bj1360259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ballard F. J. Kinetic studies with cytosol and mitochondrial phosphoenolpyruvate carboxykinases. Biochem J. 1970 Dec;120(4):809–814. doi: 10.1042/bj1200809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Berndt J., Ulbrich O. A simplified method for determination of phosphoenolpyruvate carboxykinase activity. Anal Biochem. 1970 Mar;34:282–284. doi: 10.1016/0003-2697(70)90107-7. [DOI] [PubMed] [Google Scholar]
  8. Cannata J. J. Phosphoenolpyruvate carboxykinase from bakers' yeast. Isolation of the enzyme and study of its physical properties. J Biol Chem. 1970 Feb 25;245(4):792–798. [PubMed] [Google Scholar]
  9. Chang H. C., Lane M. D. The enzymatic carboxylation of phosphoenolpyruvate. II. Purification and properties of liver mitochondrial phosphoenolpyruvate carboxykinase. J Biol Chem. 1966 May 25;241(10):2413–2420. [PubMed] [Google Scholar]
  10. Chang H. C., Maruyama H., Miller R. S., Lane M. D. The enzymatic carboxylation of phosphoenolpyruvate. 3. Investigation of the kinetics and mechanism of the mitochondrial phosphoenolpyruvate carboxykinase-catalyzed reaction. J Biol Chem. 1966 May 25;241(10):2421–2430. [PubMed] [Google Scholar]
  11. Diesterhaft M. D., Hsieh H. C., Elson C., Sallach H. J., Shrago E. Enzymatic regulation of the metabolism of phosphoenolpyruvate in Tetrahymena pyriformis. J Biol Chem. 1972 May 10;247(9):2755–2762. [PubMed] [Google Scholar]
  12. Exton J. H. Gluconeogenesis. Metabolism. 1972 Oct;21(10):945–990. doi: 10.1016/0026-0495(72)90028-5. [DOI] [PubMed] [Google Scholar]
  13. Exton J. H., Mallette L. E., Jefferson L. S., Wong E. H., Friedmann N., Miller T. B., Jr, Park C. R. The hormonal control of hepatic gluconeogenesis. Recent Prog Horm Res. 1970;26:411–461. doi: 10.1016/b978-0-12-571126-5.50014-5. [DOI] [PubMed] [Google Scholar]
  14. Felicioli R. A., Barsacchi R., Ipata P. L. Chicken liver mitochondrial phosphoenolpyruvate carboxykinase. Kinetic studies. Eur J Biochem. 1970 Apr;13(3):403–409. doi: 10.1111/j.1432-1033.1970.tb00943.x. [DOI] [PubMed] [Google Scholar]
  15. Flores H., Alleyne G. A. Phosphoenolpyruvate carboxykinase of kidney. Subcellular distribution and response to acid-base changes. Biochem J. 1971 Jun;123(1):35–39. doi: 10.1042/bj1230035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Foster D. O., Lardy H. A., Ray P. D., Johnston J. B. Alteration of rat liver phosphoenolpyruvate carboxykinase activity by L-tryptophan in vivo and metals in vitro. Biochemistry. 1967 Jul;6(7):2120–2128. doi: 10.1021/bi00859a033. [DOI] [PubMed] [Google Scholar]
  17. Foster D. O., Ray P. D., Lardy H. A. A paradoxical in vivo effect of L-tryptophan on the phosphoenolpyruvate carboxykinase of rat liver. Biochemistry. 1966 Feb;5(2):563–569. doi: 10.1021/bi00866a023. [DOI] [PubMed] [Google Scholar]
  18. Foster D. O., Ray P. D., Lardy H. A. Studies on the mechanisms underlying adaptive changes in rat liver phosphoenolpyruvate carboxykinase. Biochemistry. 1966 Feb;5(2):555–562. doi: 10.1021/bi00866a022. [DOI] [PubMed] [Google Scholar]
  19. Guder W. G., Schmidt U. The localization of gluconeogenesis in rat nephron. Determination of phosphoenolpyruvate carboxykinase in microdissected tubules. Hoppe Seylers Z Physiol Chem. 1974 Mar;355(3):273–278. doi: 10.1515/bchm2.1974.355.1.273. [DOI] [PubMed] [Google Scholar]
  20. HOLTEN D. D., NORDLIE R. C. COMPARATIVE STUDIES OF CATALYTIC PROPERTIES OF GUINEA PIG LIVER INTRA- AND EXTRAMITOCHONDRIAL PHOSPHOENOLPYRUVATE CARBOXYKINASES. Biochemistry. 1965 Apr;4:723–731. doi: 10.1021/bi00880a018. [DOI] [PubMed] [Google Scholar]
  21. Hanson R. W., Garber A. J., Reshef L., Ballard F. J. Phosphoenolpyruvate carboxykinase. II. Hormonal controls. Am J Clin Nutr. 1973 Jan;26(1):55–63. doi: 10.1093/ajcn/26.1.55. [DOI] [PubMed] [Google Scholar]
  22. Hems D. A. Metabolism of glutamine and glutamic acid by isolated perfused kidneys of normal and acidotic rats. Biochem J. 1972 Dec;130(3):671–680. doi: 10.1042/bj1300671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kamm D. E., Cahill G. F., Jr Effect of acid-base status on renal and hepatic gluconeogenesis in diabetes and fasting. Am J Physiol. 1969 May;216(5):1207–1212. doi: 10.1152/ajplegacy.1969.216.5.1207. [DOI] [PubMed] [Google Scholar]
  24. Kamm D. E., Strope G. L., Kuchmy B. L. Renal cortical and hepatic phosphoenolpyruvate carboxylase in the diabetic rat: effect of acid-base status. Metabolism. 1974 Nov;23(11):1073–1079. doi: 10.1016/0026-0495(74)90074-2. [DOI] [PubMed] [Google Scholar]
  25. Longshaw I. D., Alleyne G. A., Pogson C. I. The effect of steroids and ammonium chloride acidosis on phosphoenolpyruvate carboxykinase in rat kidney cortex. II. The kinetics of enzyme induction. J Clin Invest. 1972 Sep;51(9):2284–2291. doi: 10.1172/JCI107038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Longshaw I. D., Pogson C. I. The effect of steroids and ammonium chloride acidosis on phosphoenolpyruvate carboxykinase in rat kidney cortex. I. Differentiation of the inductive process and characterization of enzyme activities. J Clin Invest. 1972 Sep;51(9):2277–2283. doi: 10.1172/JCI107037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Martin B. R., Denton R. M. The intracellular localization of enzymes in white-adipose-tissue fat-cells and permeability properties of fat-cell mitochondria. Transfer of acetyl units and reducing power between mitochondria and cytoplasm. Biochem J. 1970 May;117(5):861–877. doi: 10.1042/bj1170861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Meyuhas O., Boshwitz C., Reshef L. Phosphoenolpyruvate carboxylase decarboxylation catalyzed reaction in cytosol of rat adipose tissue. Biochim Biophys Acta. 1971 Oct;250(1):224–237. doi: 10.1016/0005-2744(71)90138-0. [DOI] [PubMed] [Google Scholar]
  29. Murphy E. D., Anderson J. W. Tissue glycolytic and gluconeogenic enzyme activities in mildly and moderately diabetic rats: influence of tolbutamide administration. Endocrinology. 1974 Jan;94(1):27–34. doi: 10.1210/endo-94-1-27. [DOI] [PubMed] [Google Scholar]
  30. Ray P. D., Foster D. O., Lardy H. A. Paths of carbon in gluconeogenesis and lipogenesis. IV. Inhibition by L-tryptophan of hepatic gluconeogenesis at the level of phosphoenolpyruvate formation. J Biol Chem. 1966 Sep 10;241(17):3904–3908. [PubMed] [Google Scholar]
  31. Record C. O., Alberti K. G., Williamson D. H. Metabolic studies in experimental liver disease resulting from D(+)-galactosamine administration. Biochem J. 1972 Nov;130(1):37–44. doi: 10.1042/bj1300037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Reshef L., Ballard F. J., Hanson R. W. The role of the adrenals in the regulation of phosphoenolpyruvate carboxykinase of rat adipose tissue. J Biol Chem. 1969 Oct 25;244(20):5577–5581. [PubMed] [Google Scholar]
  33. Rose D. P. Aspects of tryptophan metabolism in health and disease: a review. J Clin Pathol. 1972 Jan;25(1):17–25. doi: 10.1136/jcp.25.1.17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Seubert W., Huth W. On the mechanism of gluconeogenesis and its regulation. II. The mechanism of gluconeogenesis from pyruvate and fumarate. Biochem Z. 1965 Nov 15;343(2):176–191. [PubMed] [Google Scholar]
  35. Snoke R. E., Johnston J. B., Lardy H. A. Response of phosphopyruvate carboxylase to tryptophan metabolites and metal ions. Eur J Biochem. 1971 Dec;24(2):342–346. doi: 10.1111/j.1432-1033.1971.tb19692.x. [DOI] [PubMed] [Google Scholar]
  36. Stumpf B., Boie A., Seubert W. In vitro stimulation of gluconeogenesis from pyruvate and of 14 CO 2 -fixation by dexamethasone phosphate and vasopressin in the kidney cortex of adrenalectomized rats. Biochem Biophys Res Commun. 1972 Oct 6;49(1):164–170. doi: 10.1016/0006-291x(72)90024-1. [DOI] [PubMed] [Google Scholar]
  37. Suda M., Nagai K., Nakagawa H. Studies on the circadian rhythm of phosphoenolpyruvate carboxykinase activity in rats. I. Mechanism of circadian increase in liver enzyme with special reference to hormonal and dietary effects. J Biochem. 1973 Apr;73(4):727–738. doi: 10.1093/oxfordjournals.jbchem.a130135. [DOI] [PubMed] [Google Scholar]
  38. Ui M., Claus T. H., Exton J. H., Park C. R. Studies on the mechanism of action of glucagon on gluconeogenesis. J Biol Chem. 1973 Aug 10;248(15):5344–5349. [PubMed] [Google Scholar]
  39. Walsh D. A., Chen L. J. A reinvestigation of the kinetic parameters of phosphoenolypyruvate carboxykinase. Biochem Biophys Res Commun. 1971 Nov 5;45(3):669–675. doi: 10.1016/0006-291x(71)90468-2. [DOI] [PubMed] [Google Scholar]
  40. Wicks W. D., Lewis W., McKibbin J. B. Induction of phosphoenolpyruvate carboxykinase by N 6 , O 2 '-dibutyryl cyclic AMP in rat liver. Biochim Biophys Acta. 1972 Mar 30;264(1):177–185. doi: 10.1016/0304-4165(72)90129-8. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES