Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1982 Dec 15;208(3):743–748. doi: 10.1042/bj2080743

Maximum activities of some enzymes of glycolysis, the tricarboxylic acid cycle and ketone-body and glutamine utilization pathways in lymphocytes of the rat

M Salleh M Ardawi 1, Eric A Newsholme 1
PMCID: PMC1154026  PMID: 7165729

Abstract

1. The maximum activity of hexokinase in lymphocytes is similar to that of 6-phosphofructokinase, but considerably greater than that of phosphorylase, suggesting that glucose rather than glycogen is the major carbohydrate fuel for these cells. Starvation increased slightly the activities of some of the glycolytic enzymes. A local immunological challenge in vivo (a graft-versus-host reaction) increased the activities of hexokinase, 6-phosphofructokinase, pyruvate kinase and lactate dehydrogenase, confirming the importance of the glycolytic pathway in cell division. 2. The activities of the ketone-body-utilizing enzymes were lower than those of hexokinase or 6-phosphofructokinase, unlike in muscle and brain, and were not affected by starvation. It is suggested that the ketone bodies will not provide a quantitatively important alternative fuel to glucose in lymphocytes. 3. Of the enzymes of the tricarboxylic acid cycle whose activities were measured, that of oxoglutarate dehydrogenase was the lowest, yet its activity (about 4.0μmol/min per g dry wt. at 37°C) was considerably greater than the flux through the cycle (0.5μmol/min per g calculated from oxygen consumption by incubated lymphocytes). The activity was decreased by starvation, but that of citrate synthase was increased by the local immunological challenge in vivo. It is suggested that the rate of the cycle would increase towards the capacity indicated by oxoglutarate dehydrogenase in proliferating lymphocytes. 4. Enzymes possibly involved in the pathway of glutamine oxidation were measured in lymphocytes, which suggests that an aminotransferase reaction(s) (probably aspartate aminotransferase) is important in the conversion of glutamate into oxoglutarate rather than glutamate dehydrogenase, and that the maximum activity of glutaminase is markedly in excess of the rate of glutamine utilization by incubated lymphocytes. The activity of glutaminase is increased by both starvation and the local immunological challenge in vivo. This last finding suggests that metabolism of glutamine via glutaminase is important in proliferating lymphocytes.

Full text

PDF
743

Selected References

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

  1. Alp P. R., Newsholme E. A., Zammit V. A. Activities of citrate synthase and NAD+-linked and NADP+-linked isocitrate dehydrogenase in muscle from vertebrates and invertebrates. Biochem J. 1976 Mar 15;154(3):689–700. doi: 10.1042/bj1540689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Beis A., Zammit V. A., Newsholme E. A. Activities of 3-hydroxybutyrate dehydrogenase, 3-oxoacid CoA-transferase and acetoacetyl-CoA thiolase in relation to ketone-body utilisation in muscles from vertebrates and invertebrates. Eur J Biochem. 1980 Feb;104(1):209–215. doi: 10.1111/j.1432-1033.1980.tb04417.x. [DOI] [PubMed] [Google Scholar]
  4. Boyle W. An extension of the 51Cr-release assay for the estimation of mouse cytotoxins. Transplantation. 1968 Sep;6(6):761–764. doi: 10.1097/00007890-196809000-00002. [DOI] [PubMed] [Google Scholar]
  5. Budohoski L., Challis R. A., Newsholme E. A. Effects of starvation on the maximal activities of some glycolytic and citric acid-cycle enzymes and glutaminase in mucosa of the small intestine of the rat. Biochem J. 1982 Jul 15;206(1):169–172. doi: 10.1042/bj2060169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. COOPER E. H., BARKHAN P., HALE A. J. Observations on the proliferation of human leucocytes cultured with phytohaemagglutinin. Br J Haematol. 1963 Jan;9:101–111. doi: 10.1111/j.1365-2141.1963.tb05446.x. [DOI] [PubMed] [Google Scholar]
  7. CORNBLATH M., RANDLE P. J., PARMEGGIANI A., MORGAN H. E. Regulation of glycogenolysis in muscle. Effects of glucagon and anoxia on lactate production, glycogen content, and phosphorylase activity in the perfused isolated rat heart. J Biol Chem. 1963 May;238:1592–1597. [PubMed] [Google Scholar]
  8. Crabtree B., Higgins S. J., Newsholme E. A. The activities of pyruvate carboxylase, phosphoenolpyruvate carboxylase and fructose diphosphatase in muscles from vertebrates and invertebrates. Biochem J. 1972 Nov;130(2):391–396. doi: 10.1042/bj1300391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crabtree B., Newsholme E. A. The activities of phosphorylase, hexokinase, phosphofructokinase, lactate dehydrogenase and the glycerol 3-phosphate dehydrogenases in muscles from vertebrates and invertebrates. Biochem J. 1972 Jan;126(1):49–58. doi: 10.1042/bj1260049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Culvenor J. G., Weidemann M. J. Phytohaemagglutinin stimulation of rat thymus lymphocytes glycolysis. Biochim Biophys Acta. 1976 Jul 21;437(2):354–363. doi: 10.1016/0304-4165(76)90005-2. [DOI] [PubMed] [Google Scholar]
  11. Curthoys N. P., Lowry O. H. The distribution of glutaminase isoenzymes in the various structures of the nephron in normal, acidotic, and alkalotic rat kidney. J Biol Chem. 1973 Jan 10;248(1):162–168. [PubMed] [Google Scholar]
  12. Ford W. L., Burr W., Simonsen M. A lymph node weight assay for the graft-versus-host activity of rat lymphoid cells. Transplantation. 1970 Sep;10(3):258–266. doi: 10.1097/00007890-197009000-00007. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Hanson P. J., Parsons D. S. The interrelationship between glutamine and alanine in the intestine. Biochem Soc Trans. 1980 Oct;8(5):506–509. doi: 10.1042/bst0080506. [DOI] [PubMed] [Google Scholar]
  15. Hume D. A., Radik J. L., Ferber E., Weidemann M. J. Aerobic glycolysis and lymphocyte transformation. Biochem J. 1978 Sep 15;174(3):703–709. doi: 10.1042/bj1740703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Newsholme E. A., Crabtree B., Zammit V. A. Use of enzyme activities as indices of maximum rates of fuel utilization. Ciba Found Symp. 1979;(73):245–258. doi: 10.1002/9780470720561.ch14. [DOI] [PubMed] [Google Scholar]
  17. Newsholme E. A., Lang J., Relman A. S. Control of rate of glutamine metabolism in the kidney. Contrib Nephrol. 1982;31:1–4. doi: 10.1159/000406607. [DOI] [PubMed] [Google Scholar]
  18. Newsholme E. A., Williams T. The role of phosphoenolpyruvate carboxykinase in amino acid metabolism in muscle. Biochem J. 1978 Nov 15;176(2):623–626. doi: 10.1042/bj1760623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Opie L. H., Newsholme E. A. The inhibition of skeletal-muscle fructose 1,6-diphosphatase by adenosine monophosphate. Biochem J. 1967 Aug;104(2):353–360. doi: 10.1042/bj1040353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Read G., Crabtree B., Smith G. H. The activities of 2-oxoglutarate dehydrogenase and pyruvate dehydrogenase in hearts and mammary glands from ruminants and non-ruminants. Biochem J. 1977 May 15;164(2):349–355. doi: 10.1042/bj1640349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Roos D., Loos J. A. Changes in the carbohydrate metabolism of mitogenically stimulated human peripheral lymphocytes. II. Relative importance of glycolysis and oxidative phosphorylation on phytohaemagglutinin stimulation. Exp Cell Res. 1973 Mar 15;77(1):127–135. doi: 10.1016/0014-4827(73)90561-2. [DOI] [PubMed] [Google Scholar]
  22. Ruderman N. B. Muscle amino acid metabolism and gluconeogenesis. Annu Rev Med. 1975;26:245–258. doi: 10.1146/annurev.me.26.020175.001333. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Sugden P. H., Newsholme E. A. Activities of citrate synthase, NAD+-linked and NADP+-linked isocitrate dehydrogenases, glutamate dehydrogenase, aspartate aminotransferase and alanine aminotransferase in nervous tissues from vertebrates and invertebrates. Biochem J. 1975 Jul;150(1):105–111. doi: 10.1042/bj1500105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sugden P. H., Newsholme E. A. Activities of hexokinase, phosphofructokinase, 3-oxo acid coenzyme A-transferase and acetoacetyl-coenzyme A thiolase in nervous tissue from vertebrates and invertebrates. Biochem J. 1973 May;134(1):97–101. doi: 10.1042/bj1340097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Surholt B., Newsholme E. A. Maximum activities and properties of glucose 6-phosphatase in muscles from vertebrates and invertebrates. Biochem J. 1981 Sep 15;198(3):621–629. doi: 10.1042/bj1980621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. WARBURG O. On the origin of cancer cells. Science. 1956 Feb 24;123(3191):309–314. doi: 10.1126/science.123.3191.309. [DOI] [PubMed] [Google Scholar]
  28. Watford M., Lund P., Krebs H. A. Isolation and metabolic characteristics of rat and chicken enterocytes. Biochem J. 1979 Mar 15;178(3):589–596. doi: 10.1042/bj1780589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Williamson D. H., Bates M. W., Page M. A., Krebs H. A. Activities of enzymes involved in acetoacetate utilization in adult mammalian tissues. Biochem J. 1971 Jan;121(1):41–47. doi: 10.1042/bj1210041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Williamson D. H., Lund P., Krebs H. A. The redox state of free nicotinamide-adenine dinucleotide in the cytoplasm and mitochondria of rat liver. Biochem J. 1967 May;103(2):514–527. doi: 10.1042/bj1030514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zammit V. A., Beis I., Newsholme E. A. Maximum activities and effects of fructose bisphosphate on pyruvate kinase from muscles of vertebrates and invertebrates in relation to the control of glycolysis. Biochem J. 1978 Sep 15;174(3):989–998. doi: 10.1042/bj1740989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zammit V. A., Newsholme E. A. The maximum activities of hexokinase, phosphorylase, phosphofructokinase, glycerol phosphate dehydrogenases, lactate dehydrogenase, octopine dehydrogenase, phosphoenolpyruvate carboxykinase, nucleoside diphosphatekinase, glutamate-oxaloacetate transaminase and arginine kinase in relation to carbohydrate utilization in muscles from marine invertebrates. Biochem J. 1976 Dec 15;160(3):447–462. doi: 10.1042/bj1600447. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES