Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1975 Oct;152(1):39–49. doi: 10.1042/bj1520039

Palmitate activation and esterification in microsomal fractions of rat liver.

K A Lloyd-Davies, D N Brindley
PMCID: PMC1172437  PMID: 813635

Abstract

Palmitoyl-CoA synthetase activity in the microsomal fraction of rat liver was measured directly by palmitoyl-CoA production, and indirectly by converting the palmitoyl-CoA into palmitoylcarnitine under optimum conditions. Even in the latter system, palmitoyl-CoA accumulated. The rate of palmitoyl-CoA hydrolysis and the inhibition of palmitoyl-CoA synthetase by palmitoyl-CoA were each estimated to be less than 10% of the maximum rate of palmitoyl-CoA production. The concentration of palmitoyl-CoA present in the assay systems used for measuring palmitate esterification to glycerol phosphate and the activity of palmitoyl-CoA synthetase by using the carnitine-linked determination were measured. These concentrations were not altered by the addition of glycerol phosphate, or of carnitine plus carnitine palmitoyltransferase. The relationship between the activity of palmitoyl-CoA synthetase and the rate of glycerolipid synthesis was investigated. The latter activity was measured by using palmitoyl-CoA generated from palmitate, palmitoyl--AMP or palmitoylcarnitine. It is concluded that, at optimum substrate concentrations, the activity of glycerol phosphate acyltransferase is rate-limiting in the synthesis of phosphatidate by rat liver microsomal fractions. The implications of these results in the measurement of palmitoyl-CoA synthetase and in the control of glycerolipid synthesis are discussed.

Full text

PDF
39

Selected References

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

  1. Abou-Issa H. M., Cleland W. W. Studies on the microsomal acylation of L-glycerol-3-phosphate. II. The specificity and properties of the rat liver enzyme. Biochim Biophys Acta. 1969 Jun 10;176(4):692–698. doi: 10.1016/0005-2760(69)90249-5. [DOI] [PubMed] [Google Scholar]
  2. Anderson A. D., Erwin V. G. Brain acyl-coenzyme A hydrolase: distribution, purification and properties. J Neurochem. 1971 Jul;18(7):1179–1186. doi: 10.1111/j.1471-4159.1971.tb00216.x. [DOI] [PubMed] [Google Scholar]
  3. BRANDES R., OLLEY J., SHAPIRO B. Assay of glycerol phosphate acyltransferase in liver particles. Biochem J. 1963 Feb;86:244–247. doi: 10.1042/bj0860244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bar-Tana J., Rose G., Shapiro B. The purification and properties of microsomal palmitoyl-coenzyme A synthetase. Biochem J. 1971 Apr;122(3):353–362. doi: 10.1042/bj1220353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bar-Tana J., Shapiro B. Studies on palmitoyl-coenzyme A synthetase. Biochem J. 1964 Dec;93(3):533–538. doi: 10.1042/bj0930533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Barden R. E., Cleland W. W. 1-Acylglycerol 3-phosphate acyltransferase from rat liver. J Biol Chem. 1969 Jul 10;244(13):3677–3684. [PubMed] [Google Scholar]
  7. Brandes R., Zohar Y., Arad R., Shapiro B. Two forms of microsomal palmitoyl-coenzyme A synthetase. Eur J Biochem. 1973 Apr;34(2):329–332. doi: 10.1111/j.1432-1033.1973.tb02763.x. [DOI] [PubMed] [Google Scholar]
  8. Brindley D. N., Bowley M. Drugs affecting the synthesis of glycerides and phospholipids in rat liver. The effects of clofibrate, halofenate, fenfluramine, amphetamine, cinchocaine, chlorpromazine, demethylimipramine, mepyramine and some of their derivatives. Biochem J. 1975 Jun;148(3):461–469. doi: 10.1042/bj1480461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brindley D. N., Hübscher G. The intracellular distribution of the enzymes catalysing the biosynthesis of glycerides in the intestinal mucosa. Biochim Biophys Acta. 1965 Dec 2;106(3):495–509. doi: 10.1016/0005-2760(65)90066-4. [DOI] [PubMed] [Google Scholar]
  10. Brindley D. N. The relationship between palmitoyl-coenzyme A synthetase activity and esterification of sn-glycerol 3-phosphate by the microsomal fraction of guinea-pig intestinal mucosa. Biochem J. 1973 Apr;132(4):707–715. doi: 10.1042/bj1320707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. DOLE V. P. A relation between non-esterified fatty acids in plasma and the metabolism of glucose. J Clin Invest. 1956 Feb;35(2):150–154. doi: 10.1172/JCI103259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Daae L. N., Bremer J. The acylation of glycerophosphate in rat liver. A new assay procedure for glycerophosphate acylation, studies on its subcellular and submitochondrial localization and determination of the reaction products. Biochim Biophys Acta. 1970 Jun 9;210(1):92–104. doi: 10.1016/0005-2760(70)90065-2. [DOI] [PubMed] [Google Scholar]
  13. Daae L. N. The mitochondrial acylation of glycerophosphate in rat liver: fatty acid and positional specificity. Biochim Biophys Acta. 1972 May 23;270(1):23–31. doi: 10.1016/0005-2760(72)90173-7. [DOI] [PubMed] [Google Scholar]
  14. Daniel A. M., Rubinstein D. Fatty acid esterifying enzymes in rat adipose tissue homogenates. Can J Biochem. 1968 Sep;46(9):1039–1045. doi: 10.1139/o68-156. [DOI] [PubMed] [Google Scholar]
  15. Fallon H. J., Lamb R. G. Acylation of sn-glycerol 3-phosphate by cell fractions of rat liver. J Lipid Res. 1968 Sep;9(5):652–660. [PubMed] [Google Scholar]
  16. Farstad M. In vivo and in vitro studies of the regulation of palmityl-CoA synthetase activity in rat liver. Acta Physiol Scand. 1968 Dec;74(4):568–576. doi: 10.1111/j.1748-1716.1968.tb04268.x. [DOI] [PubMed] [Google Scholar]
  17. Hajra A. K., Agranoff B. W. Acyl dihydroxyacetone phosphate. Characterization of a 32P-labeled lipid from guinea pig liver mitochondria. J Biol Chem. 1968 Apr 10;243(7):1617–1622. [PubMed] [Google Scholar]
  18. LANDS W. E., HART P. METABOLISM OF GLYCEROLIPIDS. VI. SPECIFICITIES OF ACYL COENZYME A: PHOSPHOLIPID ACYLTRANSFERASES. J Biol Chem. 1965 May;240:1905–1911. [PubMed] [Google Scholar]
  19. Lamb R. G., Fallon H. J. Inhibition of monoacylglycerophosphate formation by chlorophenoxyisobutyrate and -benzalbutyrate. J Biol Chem. 1972 Feb 25;247(4):1281–1287. [PubMed] [Google Scholar]
  20. Lamb R. G., Hill P. M., Fallon H. J. Inhibition of palmitoyl CoA deacylase by chlorophenoxyisobutyrate and betabenzalbutyrate. J Lipid Res. 1973 Jul;14(4):459–465. [PubMed] [Google Scholar]
  21. Mangiapane E. H., Lloyd-Davies K. A., Brindley D. N. A study of some enzymes of glycerolipid biosynthesis in rat liver after subtotal hepatectomy. Biochem J. 1973 May;134(1):103–112. doi: 10.1042/bj1340103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Monroy G., Kelker H. C., Pullman M. E. Partial purification and properties of an acyl coenzyme A:sn-glycerol 3-phosphate acyltransferase from rat liver mitochondria. J Biol Chem. 1973 Apr 25;248(8):2845–2852. [PubMed] [Google Scholar]
  23. NORUM K. R. PALMITYL-COA:CARNITINE PALMITYLTRANSFERASE. PURIFICATION FROM CALF-LIVER MITOCHONDRIA AND SOME PROPERTIES OF THE ENZYME. Biochim Biophys Acta. 1964 Jul 8;89:95–108. [PubMed] [Google Scholar]
  24. Pande S. V. Reversal by CoA of palmityl-CoA inhibition of long chain acyl-CoA synthetase activity. Biochim Biophys Acta. 1973 Apr 13;306(1):15–20. doi: 10.1016/0005-2760(73)90202-6. [DOI] [PubMed] [Google Scholar]
  25. Pullman M. E. A convenient and versatile method for the purification of CoA thiol esters. Anal Biochem. 1973 Jul;54(1):188–198. doi: 10.1016/0003-2697(73)90262-5. [DOI] [PubMed] [Google Scholar]
  26. Sánchez M., Nicholls D. G., Brindley D. N. [The relationship between palmitoyl-coenzyme A synthetase activity and esterification of sn-glycerol 3-phosphate in rat liver mitochondria]. Biochem J. 1973 Apr;132(4):697–706. doi: 10.1042/bj1320697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ullman M. D., Radin N. S. Oxalate-silica gel thin-layer system for free 2-hydroxy fatty acids and for fatty acyl coenzyme A. J Lipid Res. 1972 May;13(3):422–423. [PubMed] [Google Scholar]
  28. VIGNAIS P. V., ZABIN I. Synthesis and properties of palmityl adenylate, palmityl coenzyme A, and palmityl glutathione. Biochim Biophys Acta. 1958 Aug;29(2):263–269. doi: 10.1016/0006-3002(58)90183-5. [DOI] [PubMed] [Google Scholar]
  29. Yamashita S., Numa S. Partial purification and properties of glycerophosphate acyltransferase from rat liver. Formation of 1-acylglycerol 3-phosphate from sn-glycerol 3-phosphate and palmityl coenzyme A. Eur J Biochem. 1972 Dec 18;31(3):565–573. doi: 10.1111/j.1432-1033.1972.tb02566.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES