Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1968 Nov;110(1):27–38. doi: 10.1042/bj1100027

The control of fatty acid and triglyceride synthesis in rat epididymal adipose tissue. Roles of coenzyme A derivatives, citrate and l-glycerol 3-phosphate

R M Denton 1, M L Halperin 1,*
PMCID: PMC1187105  PMID: 5749224

Abstract

1. Methods are described for the extraction and assay of acetyl-CoA and of total acid-soluble and total acid-insoluble CoA derivatives in rat epididymal adipose tissue. 2. The concentration ranges of the CoA derivatives in fat pads incubated in vitro under various conditions were: total acid-soluble CoA, 0·20–0·59mm; total acid-insoluble CoA, 0·08–0·23mm; acetyl-CoA, 0·03–0·14mm. 3. An investigation was made of some postulated mechanisms of control of fatty acid and triglyceride synthesis in rat epididymal fat pads incubated in vitro. The concentrations of intermediates of possible regulatory significance were measured at various rates of fatty acid and triglyceride synthesis produced by the addition to the incubation medium (Krebs bicarbonate buffer containing glucose) of insulin, adrenaline, albumin, palmitate or acetate. 4. The whole-tissue concentrations of glucose 6-phosphate, l-glycerol 3-phosphate, citrate, acetyl-CoA, total acid-soluble CoA and total acid-insoluble CoA were assayed after 30 or 60min. incubation. The rates of fatty acid and triglyceride synthesis, calculated from the incorporation of [U-14C]glucose into fatty acids and glyceride glycerol respectively, and the rates of glucose uptake, lactate plus pyruvate output and glycerol output were measured over a 60min. incubation. 5. The rate of triglyceride synthesis could not be correlated with the concentrations of either l-glycerol 3-phosphate or long-chain fatty acyl-CoA (measured as total acid-insoluble CoA). Factor(s) other than the whole-tissue concentrations of these recognized precursors appear to be involved in the determination of the rate of triglyceride synthesis. 6. No relationship was found between the rate of fatty acid synthesis and the whole-tissue concentrations of the intermediates, citrate or acetyl-CoA, or with the two proposed effectors of acetyl-CoA carboxylase, citrate (as activator) or long-chain fatty acyl-CoA (as inhibitor). The control of fatty acid synthesis appears to reside in additional or alternative factors.

Full text

PDF
27

Selected References

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

  1. BALL E. G., MARTIN D. B., COOPER O. Studies on the metabolism of adipose tissue. I. The effect of insulin on glucose utilization as measured by the manometric determination of carbon dioxide output. J Biol Chem. 1959 Apr;234(4):774–780. [PubMed] [Google Scholar]
  2. BALLY P. R., CAHILL G. F., Jr, LEBOEUF B., RENOLD A. E. Studies on rat adipose tissue in vitro. V. Effects of glucose and insulin on the metabolism of palmitate-1-C14. J Biol Chem. 1960 Feb;235:333–336. [PubMed] [Google Scholar]
  3. BORTZ W. M., LYNEN F. THE INHIBITION OF ACETYL COA CARBOXYLASE BY LONG CHAIN ACYL COA DERIVATIVES. Biochem Z. 1963 Aug 14;337:505–509. [PubMed] [Google Scholar]
  4. CAHILL G. F., Jr, LEBOEUF B., FLINN R. B. Studies on rat adipose tissue in vitro. VI. Effect of epinephrine on glucose metabolism. J Biol Chem. 1960 May;235:1246–1250. [PubMed] [Google Scholar]
  5. CROFFORD O. B., RENOLD A. E. GLUCOSE UPTAKE BY INCUBATED RAT EPIDIDYMAL ADIPOSE TISSUE. CHARACTERISTICS OF THE GLUCOSE TRANSPORT SYSTEM AND ACTION OF INSULIN. J Biol Chem. 1965 Aug;240:3237–3244. [PubMed] [Google Scholar]
  6. CROFFORD O. B., RENOLD A. E. GLUCOSE UPTAKE BY INCUBATED RAT EPIDIDYMAL ADIPOSE TISSUE. RATE-LIMITING STEPS AND SITE OF INSULIN ACTION. J Biol Chem. 1965 Jan;240:14–21. [PubMed] [Google Scholar]
  7. Chase J. F. pH-dependence of carnitine acetyltransferase activity. Biochem J. 1967 Aug;104(2):503–509. doi: 10.1042/bj1040503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chen R. F. Removal of fatty acids from serum albumin by charcoal treatment. J Biol Chem. 1967 Jan 25;242(2):173–181. [PubMed] [Google Scholar]
  9. DOLE V. P., MEINERTZ H. Microdetermination of long-chain fatty acids in plasma and tissues. J Biol Chem. 1960 Sep;235:2595–2599. [PubMed] [Google Scholar]
  10. Denton R. M., Randle P. J. Citrate and the regulation of adipose-tissue phosphofructokinase. Biochem J. 1966 Aug;100(2):420–423. doi: 10.1042/bj1000420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Denton R. M., Randle P. J. Measurement of flow of carbon atoms from glucose and glycogen glucose to glyceride glycerol and glycerol in rat heart and epididymal adipose tissue. Effects of insulin, adrenaline and alloxan-diabetes. Biochem J. 1967 Aug;104(2):423–434. doi: 10.1042/bj1040423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Denton R. M., Yorke R. E., Randle P. J. Measurement of concentrations of metabolites in adipose tissue and effects of insulin, alloxan-diabetes and adrenaline. Biochem J. 1966 Aug;100(2):407–419. doi: 10.1042/bj1000407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Duncombe W. G. The colorimetric micro-determination of long-chain fatty acids. Biochem J. 1963 Jul;88(1):7–10. doi: 10.1042/bj0880007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. FLATT J. P., BALL E. G. STUDIES ON THE METABOLISM OF ADIPOSE TISSUE. XV. AN EVALUATION OF THE MAJOR PATHWAYS OF GLUCOSE CATABOLISM AS INFLUENCED BY INSULIN AND EPINEPHRINE. J Biol Chem. 1964 Mar;239:675–685. [PubMed] [Google Scholar]
  15. Fang M., Lowenstein J. M. Citrate and the conversion of carbohydrate into fat. The regulation of fatty acid synthesis by rat liver extracts. Biochem J. 1967 Nov;105(2):803–811. doi: 10.1042/bj1050803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Flatt J. P., Ball E. G. Studies on the metabolism of adipose tissue. XIX. An evaluation of the major pathways of glucose catabolism as influenced by acetate in the presence of insulin. J Biol Chem. 1966 Jun 25;241(12):2862–2869. [PubMed] [Google Scholar]
  17. GARLAND P. B., RANDLE P. J. A rapid enzymatic assay for glycerol. Nature. 1962 Dec 8;196:987–988. doi: 10.1038/196987a0. [DOI] [PubMed] [Google Scholar]
  18. Garland P. B., Newsholme E. A., Randle P. J. Regulation of glucose uptake by muscle. 9. Effects of fatty acids and ketone bodies, and of alloxan-diabetes and starvation, on pyruvate metabolism and on lactate-pyruvate and L-glycerol 3-phosphate-dihydroxyacetone phosphate concentration ratios in rat heart and rat diaphragm muscles. Biochem J. 1964 Dec;93(3):665–678. doi: 10.1042/bj0930665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Garland P. B., Shepherd D., Yates D. W. Steady-state concentrations of coenzyme A, acetyl-coenzyme A and long-chain fatty acyl-coenzyme A in rat-liver mitochondria oxidizing palmitate. Biochem J. 1965 Nov;97(2):587–594. doi: 10.1042/bj0970587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. LANDAU B. R., BARTSCH G. E., KATZ J., WOOD H. G. ESTIMATION OF PATHWAY CONTRIBUTIONS TO GLUCOSE METABOLISM AND OF THE RATE OF ISOMERIZATION OF HEXOSE 6-PHOSPHATE. J Biol Chem. 1964 Mar;239:686–696. [PubMed] [Google Scholar]
  21. MARQUIS N. R., FRITZ I. B. ENZYMOLOGICAL DETERMINATION OF FREE CARNITINE CONCENTRATIONS IN RAT TISSUES. J Lipid Res. 1964 Apr;5:184–187. [PubMed] [Google Scholar]
  22. MARTIN D. B., VAGELOS P. R. The mechanism of tricarboxylic acid cycle regulation of fatty acid synthesis. J Biol Chem. 1962 Jun;237:1787–1792. [PubMed] [Google Scholar]
  23. Moellering H., Gruber W. Determination of citrate with citrate lyase. Anal Biochem. 1966 Dec;17(3):369–376. doi: 10.1016/0003-2697(66)90172-2. [DOI] [PubMed] [Google Scholar]
  24. Pearson D. J., Tubbs P. K. Carnitine and derivatives in rat tissues. Biochem J. 1967 Dec;105(3):953–963. doi: 10.1042/bj1050953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Randle P. J., Denton R. M., England P. J. Citrate as a metabolic regulator in muscle and adipose tissue. Biochem Soc Symp. 1968;27:87–103. [PubMed] [Google Scholar]
  26. Randle P. J., Newsholme E. A., Garland P. B. Regulation of glucose uptake by muscle. 8. Effects of fatty acids, ketone bodies and pyruvate, and of alloxan-diabetes and starvation, on the uptake and metabolic fate of glucose in rat heart and diaphragm muscles. Biochem J. 1964 Dec;93(3):652–665. doi: 10.1042/bj0930652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rao G. A., Johnston J. M. Studies of the formation and utilization of bound CoA in glyceride biosynthesis. Biochim Biophys Acta. 1967 Aug 8;144(1):25–33. doi: 10.1016/0005-2760(67)90074-4. [DOI] [PubMed] [Google Scholar]
  28. SANADI D. R., LITTLEFIELD J. W., BOCK R. M. Studies on alpha-ketoglutaric oxidase. II. Purification and properties. J Biol Chem. 1952 May;197(2):851–862. [PubMed] [Google Scholar]
  29. SHORT D. J., PARKES A. S. Feeding and breeding of laboratory animals; a compound diet for monkeys. J Hyg (Lond) 1949 Jun;47(2):209–212. doi: 10.1017/s0022172400014480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. SMITH S. W., WEISS S. B., KENNEDY E. P. The enzymatic dephosphorylation of phosphatidic acids. J Biol Chem. 1957 Oct;228(2):915–922. [PubMed] [Google Scholar]
  31. TROUT D. L., ESTES E. H., Jr, FRIEDBERG S. J. Titration of free fatty acids of plasma: a study of current methods and a new modification. J Lipid Res. 1960 Apr;1:199–202. [PubMed] [Google Scholar]
  32. Tubbs P. K., Garland P. B. Variations in tissue contents of coenzyme A thio esters and possible metabolic implications. Biochem J. 1964 Dec;93(3):550–557. doi: 10.1042/bj0930550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. VAGELOS P. R., ALBERTS A. W., MARTIN D. B. Studies on the mechnism of activation of acetyl coenzyme A carboxylase by citrate. J Biol Chem. 1963 Feb;238:533–540. [PubMed] [Google Scholar]
  34. VALLANCE-OWEN J., DENNES E., CAMPBELL P. N. Insulin antagonism in plasma of diabetic patients and normal subjects. Lancet. 1958 Aug 16;2(7042):336–338. doi: 10.1016/s0140-6736(58)90257-5. [DOI] [PubMed] [Google Scholar]
  35. WIELAND O., WEISS L. INHIBITION OF CITRATE-SYNTHASE BY PALMITYL-COENZYME A. Biochem Biophys Res Commun. 1963 Sep 10;13:26–31. doi: 10.1016/0006-291x(63)90156-6. [DOI] [PubMed] [Google Scholar]
  36. WINEGRAD A. I., RENOLD A. E. Studies on rat adipose tissue in vitro. I. Effects of insulin on the metabolism of glucose, pyruvate, and acetate. J Biol Chem. 1958 Aug;233(2):267–272. [PubMed] [Google Scholar]
  37. 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]

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

RESOURCES