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. 1991 Feb 15;274(Pt 1):145–152. doi: 10.1042/bj2740145

Identity between palmitoyl-CoA synthetase and arachidonoyl-CoA synthetase in human platelet?

A M Bakken 1, M Farstad 1, H Holmsen 1
PMCID: PMC1149932  PMID: 1848073

Abstract

Apparent Km values have been determined for the substrates ATP, CoA and fatty acids for the long-chain acyl-CoA synthetase (EC 6.2.1.3) reaction in lysates of human blood platelets. The apparent Km for ATP was higher for saturated fatty acids (C12:0 to C18:0) than for unsaturated acids (C18:1 to C22:6). Other apparent Km values were very similar for all long-chain fatty acids tested. Palmitic acid inhibited the formation of [14C]arachidonoyl-CoA, and arachidonic acid inhibited the formation of [14C]palmitoyl-CoA, with [14C]arachidonate or [14C]palmitate respectively as substrate. After chromatography of Triton X-100-extracted platelet protein in several systems (hydroxyapatite, DEAE-Sepharose, Sephacryl S-200 HR, CoA-Sepharose, Sephadex G-100 and AcA 34), both arachidonoyl-CoA synthetase and palmitoyl-CoA synthetase activities were eluted together in the various protein peaks, and with approximately the same ratio of activities in all peaks. After some purification steps (DEAE-Sepharose and Sephacryl S-200 HR), the acyl-CoA synthetase activity was up to 37 nmol/min per mg of protein with [14C]palmitate as substrate, and up to 116 nmol/min per mg of protein with [14C]arachidonate as substrate. The purification was respectively about 8- and 10-fold. The results indicate that palmitoyl-CoA (or unspecific) synthetase and arachidonoyl-CoA (or specific) synthetase are in fact the same enzyme, in agreement with previously reported results from this laboratory.

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Selected References

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  1. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  2. Bakken A. M., Farstad M. Identical subcellular distribution of palmitoyl-CoA and arachidonoyl-CoA synthetase activities in human blood platelets. Biochem J. 1989 Jul 1;261(1):71–76. doi: 10.1042/bj2610071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Berge R. K., Farstad M. Hydrolysis of long-chain fatty acyl-CoA in homogenates of human blood platelets: the existence of a platelet palmitoyl-CoA hydrolase. Scand J Clin Lab Invest. 1978 Dec;38(8):699–706. doi: 10.1080/00365517809104876. [DOI] [PubMed] [Google Scholar]
  5. Berge R. K., Slinde E., Farstad M. Variations in the activity of microsomal palmitoyl-CoA hydrolase in mixed micelle solutions of palmitoyl-CoA and non-ionic detergents of the triton X series. Biochim Biophys Acta. 1981 Oct 23;666(1):25–35. doi: 10.1016/0005-2760(81)90087-4. [DOI] [PubMed] [Google Scholar]
  6. Berge R. K., Vollset S. E., Farstad M. Intracellular localization of palmitoyl-CoA hydrolase and palmitoyl-CoA synthetase in human blood platelets and liver. Scand J Clin Lab Invest. 1980 May;40(3):271–278. doi: 10.3109/00365518009095578. [DOI] [PubMed] [Google Scholar]
  7. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  8. Farstad M., Aas M., Sander J. Long-chain acyl-CoA synthetase activity in blood platelets and some human tissues. Scand J Clin Lab Invest. 1973 Mar;31(2):205–211. doi: 10.3109/00365517309084311. [DOI] [PubMed] [Google Scholar]
  9. Farstad M., Sander J. The existence of a long-chain acyl-CoA synthetase in homogenates of human blood platelets. Scand J Clin Lab Invest. 1971 Nov;28(3):261–265. doi: 10.3109/00365517109095698. [DOI] [PubMed] [Google Scholar]
  10. Hauser G., Eichberg J. Identification of cytidine diphosphate-diglyceride in the pineal gland of the rat and its accumulation in the presence of DL-propranolol. J Biol Chem. 1975 Jan 10;250(1):105–112. [PubMed] [Google Scholar]
  11. Holmsen H., Ostvold A. C., Pimentel M. A. Enzymatic properties of 5'-AMP deaminase in platelet lysates. Thromb Haemost. 1977 Jun 30;37(3):380–395. [PubMed] [Google Scholar]
  12. Ingebretsen O. C., Bakken A. M., Farstad M. The content of coenzyme A, acetyl-CoA and long-chain acyl-CoA in human blood platelets. Clin Chim Acta. 1982 Dec 23;126(3):307–313. doi: 10.1016/0009-8981(82)90305-9. [DOI] [PubMed] [Google Scholar]
  13. Ingebretsen O. C., Bakken A. M., Segadal L., Farstad M. Determination of adenine nucleotides and inosine in human myocard by ion-pair reversed-phase high-performance liquid chromatography. J Chromatogr. 1982 Jun 18;242(1):119–126. doi: 10.1016/s0021-9673(00)87253-2. [DOI] [PubMed] [Google Scholar]
  14. Iritani N., Ikeda Y., Kajitani H. Selectivities of 1-acylglycerophosphorylcholine acyltransferase and acyl-CoA synthetase for n-3 polyunsaturated fatty acids in platelets and liver microsomes. Biochim Biophys Acta. 1984 May 11;793(3):416–422. [PubMed] [Google Scholar]
  15. Kameda K., Suzuki L. K., Imai Y. Further purification, characterization and salt activation of acyl-CoA synthetase from Escherichia coli. Biochim Biophys Acta. 1985 May 29;840(1):29–36. doi: 10.1016/0304-4165(85)90158-8. [DOI] [PubMed] [Google Scholar]
  16. Kramer R. M., Pritzker C. R., Deykin D. Coenzyme A-mediated arachidonic acid transacylation in human platelets. J Biol Chem. 1984 Feb 25;259(4):2403–2406. [PubMed] [Google Scholar]
  17. Laposata M., Krueger C. M., Saffitz J. E. Selective uptake of [3H]arachidonic acid into the dense tubular system of human platelets. Blood. 1987 Sep;70(3):832–837. [PubMed] [Google Scholar]
  18. Laposata M., Reich E. L., Majerus P. W. Arachidonoyl-CoA synthetase. Separation from nonspecific acyl-CoA synthetase and distribution in various cells and tissues. J Biol Chem. 1985 Sep 15;260(20):11016–11020. [PubMed] [Google Scholar]
  19. Lienhard G. E., Secemski I. I. P 1 ,P 5 -Di(adenosine-5')pentaphosphate, a potent multisubstrate inhibitor of adenylate kinase. J Biol Chem. 1973 Feb 10;248(3):1121–1123. [PubMed] [Google Scholar]
  20. Majerus P. W., Prescott S. M., Hofmann S. L., Neufeld E. J., Wilson D. B. Uptake and release of arachidonate by platelets. Adv Prostaglandin Thromboxane Leukot Res. 1983;11:45–52. [PubMed] [Google Scholar]
  21. McKean M. L., Smith J. B., Silver M. J. Phospholipid biosynthesis in human platelets. Formation of phosphatidylcholine from 1-acyl lysophosphatidylcholine by acyl-CoA:1-acyl-sn-glycero-3-phosphocholine acyltransferase. J Biol Chem. 1982 Oct 10;257(19):11278–11283. [PubMed] [Google Scholar]
  22. Neufeld E. J., Sprecher H., Evans R. W., Majerus P. W. Fatty acid structural requirements for activity of arachidonoyl-CoA synthetase. J Lipid Res. 1984 Mar;25(3):288–293. [PubMed] [Google Scholar]
  23. Norum K. R., Farstad M., Bremer J. The submitochondrial distribution of acid:CoA ligase (AMP) and palmityl-CoA:carnitine palmityltransferase in rat liver mitochondria. Biochem Biophys Res Commun. 1966 Sep 8;24(5):797–804. doi: 10.1016/0006-291x(66)90397-4. [DOI] [PubMed] [Google Scholar]
  24. Noy N., Zakim D. Fatty acids bound to unilamellar lipid vesicles as substrates for microsomal acyl-CoA ligase. Biochemistry. 1985 Jul 2;24(14):3521–3525. doi: 10.1021/bi00335a020. [DOI] [PubMed] [Google Scholar]
  25. Ockner R. K., Manning J. A. Fatty acid-binding protein in small intestine. Identification, isolation, and evidence for its role in cellular fatty acid transport. J Clin Invest. 1974 Aug;54(2):326–338. doi: 10.1172/JCI107768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pande S. V., Lee T. S., Murthy M. S. Freeze-thawing causes masking of membrane-bound outer carnitine palmitoyltransferase activity: implications for studies on carnitine palmitoyltransferases deficiency. Biochim Biophys Acta. 1990 May 22;1044(2):262–268. doi: 10.1016/0005-2760(90)90312-l. [DOI] [PubMed] [Google Scholar]
  27. Suzuki H., Kawarabayasi Y., Kondo J., Abe T., Nishikawa K., Kimura S., Hashimoto T., Yamamoto T. Structure and regulation of rat long-chain acyl-CoA synthetase. J Biol Chem. 1990 May 25;265(15):8681–8685. [PubMed] [Google Scholar]
  28. Tanaka T., Hosaka K., Hoshimaru M., Numa S. Purification and properties of long-chain acyl-coenzyme-A synthetase from rat liver. Eur J Biochem. 1979 Jul;98(1):165–172. doi: 10.1111/j.1432-1033.1979.tb13173.x. [DOI] [PubMed] [Google Scholar]
  29. Vollset S. E., Farstad M. A study of assay conditions for palmitoyl-CoA synthetase and carnitine palmitoyltransferase in homogenates of human blood platelets. Scand J Clin Lab Invest. 1979 Feb;39(1):15–21. doi: 10.3109/00365517909104934. [DOI] [PubMed] [Google Scholar]
  30. Wilson D. B., Prescott S. M., Majerus P. W. Discovery of an arachidonoyl coenzyme A synthetase in human platelets. J Biol Chem. 1982 Apr 10;257(7):3510–3515. [PubMed] [Google Scholar]

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