Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1976 Sep 15;158(3):575–581. doi: 10.1042/bj1580575

Incorporation of choline and ethanolamine into phospholipids in germinating soya bean.

C W Dykes, J Kay, J L Harwood
PMCID: PMC1164012  PMID: 988830

Abstract

1. Incorporation of [Me-14C]choline and [2-14C]ethanolamine into lipids was studied in germinating soya bean (Glycine max L.) seeds. The precursors are only incorporated into phosphatidylcholine and into phosphatidylethanolamine respectively. 2. Base-labelling via a phospholipase-D type of reaction was eliminated as a significant factor. 3. Cyclo heximide inhibited labelling of phosphatidylcholine from [Me-14C]choline but did not affect labelling of the aqueous choline pool. It had no effect on [2-14C]ethanolamine uptake or incorporation into phosphatidylethanolamine. 4. Hemicholinium-15 at 10mM concentrations decreased uptake and lipid labelling from the both bases. 5. There was no evidence for base competition. 6. The endogenous pool of choline was much larger than that of ethanolamine, which resulted in higher specific radioactivities for phosphatidyl-ethanolamine than for phosphatidylcholine. 7. The results can be interpreted as indicating that the kinase and phosphoryltransferase enzymes of the CDP-base pathways are separate for each phospholipid.

Full text

PDF
576

Selected References

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

  1. APP A. A., JAGENDORF A. T. INCORPORATION OF LABELLED AMINO ACIDS BY CHLOROPLAST RIBOSOMES. Biochim Biophys Acta. 1963 Oct 15;76:286–292. [PubMed] [Google Scholar]
  2. Abdelkader A. B., Cherif A., Demandre C., Mazliak P. The oleyl-coenzyme-A desaturase of potato tubers. Enzymatic properties, intracellular localization and induction during "aging" of tuber slices. Eur J Biochem. 1973 Jan 3;32(1):155–165. doi: 10.1111/j.1432-1033.1973.tb02592.x. [DOI] [PubMed] [Google Scholar]
  3. Ansell G. B., Spanner S. G. The inhibition of brain choline kinase by hemicholinium-3. J Neurochem. 1974 Jun;22(6):1153–1155. doi: 10.1111/j.1471-4159.1974.tb04352.x. [DOI] [PubMed] [Google Scholar]
  4. Broad T. E., Dawson R. M. Phospholipid biosynthesis in the anaerobic protozoon Entodinium caudatum. Biochem J. 1975 Feb;146(2):317–328. doi: 10.1042/bj1460317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brockerhoff H., Yurkowski M. Stereospecific analyses of several vegetable fats. J Lipid Res. 1966 Jan;7(1):62–64. [PubMed] [Google Scholar]
  6. Bygrave F. L., Dawson R. M. Phosphatidylcholine biosynthesis in the anaerobic protozoon Entodinium caudatum. Biochem Soc Trans. 1975;3(5):740–741. doi: 10.1042/bst0030740. [DOI] [PubMed] [Google Scholar]
  7. Dawson R. M. The formation of phosphatidylglycerol and other phospholipids by the transferase activity of phospholipase D. Biochem J. 1967 Jan;102(1):205–210. doi: 10.1042/bj1020205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Devor K. A., Mudd J. B. Biosynthesis of phosphatidylcholine by enzyme preparations from spinach leaves. J Lipid Res. 1971 Jul;12(4):403–411. [PubMed] [Google Scholar]
  9. GARBUS J., DELUCA H. F., LOOMANS M. E., STRONG F. M. The rapid incorporation of phosphate into mitochondrial lipids. J Biol Chem. 1963 Jan;238:59–63. [PubMed] [Google Scholar]
  10. Gurr M. I., Robinson M. P., James A. T. The mechanism of formation of polyunsaturated fatty acids by photosynthetic tissue. The tight coupling of oleate desaturation with phospholipid synthesis in Chlorella vulgaris. Eur J Biochem. 1969 May 1;9(1):70–78. doi: 10.1111/j.1432-1033.1969.tb00577.x. [DOI] [PubMed] [Google Scholar]
  11. Harwood J. L. Biosynthesis of phosphatidylcholine and phosphatidylethanolamine by germinating soya bean. Biochem Soc Trans. 1976;4(1):50–52. doi: 10.1042/bst0040050. [DOI] [PubMed] [Google Scholar]
  12. Harwood J. L., Desai R., Hext P., Tetley T., Richards R. Characterization of pulmonary surfactant from ox, rabbit, rat and sheep. Biochem J. 1975 Dec;151(3):707–714. doi: 10.1042/bj1510707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hemsworth B. A., Darmer K. I., Jr, Bosmann H. B. The incorporation of choline into isolated synaptosomal and synaptic vesicle fractions in the presence of quaternary ammonium compounds. Neuropharmacology. 1971 Jan;10(1):109–119. doi: 10.1016/0028-3908(71)90014-1. [DOI] [PubMed] [Google Scholar]
  14. Hemsworth B. A. The effect of a hemicholinium analogue, HC-15, of neuromuscular transmission. Eur J Pharmacol. 1971 Jun;15(1):91–100. doi: 10.1016/0014-2999(71)90083-5. [DOI] [PubMed] [Google Scholar]
  15. Hoelzl J., Wagner H. Simultaneous P32- and C14-labeling of phospholipids by germinating soybeans. J Lipid Res. 1966 Jul;7(4):569–570. [PubMed] [Google Scholar]
  16. Hölzl J., Wagner H. Uber eine in-vivo-Methode zur Herstellung radioaktiv markerter Phosphatide mit Sojabohnen. Z Naturforsch B. 1971 May;26(5):425–434. [PubMed] [Google Scholar]
  17. Lord J. M. Evidence that phosphatidylcholine and phosphatidylethanolamine are synthesized by a single enzyme present in the endoplasmic reticulum of castor-bean endosperm. Biochem J. 1975 Nov;151(2):451–453. doi: 10.1042/bj1510451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lord J. M., Kagawa T., Beevers H. Intracellular distribution of enzymes of the cytidine diphosphate choline pathway in castor bean endosperm. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2429–2432. doi: 10.1073/pnas.69.9.2429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marcus A., Feeley J. Ribosome activation and polysome formation in vitro: requirement for ATP. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1770–1777. doi: 10.1073/pnas.56.6.1770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marshall M. O., Kates M. Biosynthesis of nitrogenous phospholipids in spinach leaves. Can J Biochem. 1974 Jun;52(6):469–482. doi: 10.1139/o74-071. [DOI] [PubMed] [Google Scholar]
  21. Miedema E., Richardson K. E. Ethanolamine metabolism in plant tissues. Plant Physiol. 1966 Jun;41(6):1026–1030. doi: 10.1104/pp.41.6.1026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Moore T. S., Lord J. M., Kagawa T., Beevers H. Enzymes of phospholipid metabolism in the endoplasmic reticulum of castor bean endosperm. Plant Physiol. 1973 Jul;52(1):50–53. doi: 10.1104/pp.52.1.50. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Moore T. S. Phosphatidylcholine synthesis in castor bean endosperm. Plant Physiol. 1976 Mar;57(3):382–386. doi: 10.1104/pp.57.3.382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Morré D. J., Nyquist S., Rivera E. Lecithin Biosynthetic Enzymes of Onion Stem and the Distribution of Phosphorylcholine-Cytidyl Transferase among Cell Fractions. Plant Physiol. 1970 Jun;45(6):800–804. doi: 10.1104/pp.45.6.800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Privett O. S., Dougherty K. A., Erdahl W. L., Stolyhwo A. Studies on the lipid composition of developing soybeans. J Am Oil Chem Soc. 1973 Dec;50(12):516–520. doi: 10.1007/BF02640523. [DOI] [PubMed] [Google Scholar]
  26. Pugh E. L., Kates M. Desaturation of phosphatidylcholine and phosphatidylethanolamine by a microsomal enzyme system from Candida lipolytica. Biochim Biophys Acta. 1973 Sep 25;316(3):305–316. doi: 10.1016/0005-2760(73)90071-4. [DOI] [PubMed] [Google Scholar]
  27. RAMASARMA T., WETTER L. R. Choline kinase of rapeseed (Brassica campestris L.). Can J Biochem Physiol. 1957 Oct;35(10):853–863. [PubMed] [Google Scholar]
  28. Roughan P. G., Slack C. R. Is phospholipase D really an enzyme? A comparison of in situ and in vitro activities. Biochim Biophys Acta. 1976 Apr 22;431(1):86–95. doi: 10.1016/0005-2760(76)90262-9. [DOI] [PubMed] [Google Scholar]
  29. Roughan P. G. Turnover of the glycerolipids of pumpkin leaves. The importence of phosphatidylcholine. Biochem J. 1970 Mar;117(1):1–8. doi: 10.1042/bj1170001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tanaka K., Tolbert N. E., Gohlke A. F. Choline kinase and phosphorylcholine phosphatase in plants. Plant Physiol. 1966 Feb;41(2):307–312. doi: 10.1104/pp.41.2.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tang W. J., Castelfranco P. A. Phospholipid synthesis in aging potato tuber tissue. Plant Physiol. 1968 Aug;43(8):1232–1238. doi: 10.1104/pp.43.8.1232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vandor S. L., Richardson K. E. Incorporation of ethanolamine-1,2-14C into plant microsomal phospholipids. Can J Biochem. 1968 Oct;46(10):1309–1315. doi: 10.1139/o68-196. [DOI] [PubMed] [Google Scholar]
  33. Vijay I. K., Stumpf P. K. Fat metabolism in higher plants. XLVI. Nature of the substrate and the product of oleyl coenzyme A desaturase from Carthamus tinctorius. J Biol Chem. 1971 May 10;246(9):2910–2917. [PubMed] [Google Scholar]
  34. Willemot C., Verret G. Incorporation of choline-1,2-14C into molecular species of phosphatidylcholine by alfalfa leaflet tissue. Lipids. 1973 Oct;8(10):588–591. doi: 10.1007/BF02532716. [DOI] [PubMed] [Google Scholar]
  35. Yang S. F., Freer S., Benson A. A. Transphosphatidylation by phospholipase D. J Biol Chem. 1967 Feb 10;242(3):477–484. [PubMed] [Google Scholar]

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

RESOURCES