Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1989 Oct 15;263(2):581–587. doi: 10.1042/bj2630581

Analysis of the water-soluble products of phosphatidylcholine breakdown by ion-exchange chromatography. Bombesin and TPA (12-O-tetradecanoylphorbol 13-acetate) stimulate choline generation in Swiss 3T3 cells by a common mechanism.

S J Cook 1, M J Wakelam 1
PMCID: PMC1133466  PMID: 2597122

Abstract

A method for the rapid and quantitative separation of glycerophosphocholine, choline phosphate and choline upon ion-exchange columns is described. The method has been utilized to examine the stimulation of phosphatidylcholine breakdown in quiescent Swiss 3T3 cells in response to bombesin and 12-O-tetradecanoylphorbol 13-acetate (TPA). The stimulated generation of choline is shown to precede that of choline phosphate, with no effect upon glycerophosphocholine levels; but was attenuated in cells in which protein kinase C activity was down-regulated. The results thus suggest that stimulation of the cells with either bombesin or TPA activates phospholipase D-catalysed phosphatidylcholine breakdown by a common mechanism involving the activation of protein kinase C.

Full text

PDF
584

Selected References

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

  1. Berridge M. J. Inositol lipids and cell proliferation. Biochim Biophys Acta. 1987 Apr 20;907(1):33–45. doi: 10.1016/0304-419x(87)90017-5. [DOI] [PubMed] [Google Scholar]
  2. Bocckino S. B., Blackmore P. F., Wilson P. B., Exton J. H. Phosphatidate accumulation in hormone-treated hepatocytes via a phospholipase D mechanism. J Biol Chem. 1987 Nov 5;262(31):15309–15315. [PubMed] [Google Scholar]
  3. Brown K. D., Blakeley D. M., Hamon M. H., Laurie M. S., Corps A. N. Protein kinase C-mediated negative-feedback inhibition of unstimulated and bombesin-stimulated polyphosphoinositide hydrolysis in Swiss-mouse 3T3 cells. Biochem J. 1987 Aug 1;245(3):631–639. doi: 10.1042/bj2450631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown K. D., Blay J., Irvine R. F., Heslop J. P., Berridge M. J. Reduction of epidermal growth factor receptor affinity by heterologous ligands: evidence for a mechanism involving the breakdown of phosphoinositides and the activation of protein kinase C. Biochem Biophys Res Commun. 1984 Aug 30;123(1):377–384. doi: 10.1016/0006-291x(84)90424-8. [DOI] [PubMed] [Google Scholar]
  5. Cabot M. C., Welsh C. J., Cao H. T., Chabbott H. The phosphatidylcholine pathway of diacylglycerol formation stimulated by phorbol diesters occurs via phospholipase D activation. FEBS Lett. 1988 Jun 6;233(1):153–157. doi: 10.1016/0014-5793(88)81374-7. [DOI] [PubMed] [Google Scholar]
  6. Castagna M., Takai Y., Kaibuchi K., Sano K., Kikkawa U., Nishizuka Y. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem. 1982 Jul 10;257(13):7847–7851. [PubMed] [Google Scholar]
  7. Clarke N. G., Dawson R. M. Alkaline O leads to N-transacylation. A new method for the quantitative deacylation of phospholipids. Biochem J. 1981 Apr 1;195(1):301–306. doi: 10.1042/bj1950301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dowdall M. J., Barker L. A., Whittaker V. P. Choline metabolism in the cerebral cortex of guinea pigs. Phosphorylcholine and lipid choline. Biochem J. 1972 Dec;130(4):1081–1094. doi: 10.1042/bj1301081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gardner S. D., Milligan G., Rice J. E., Wakelam M. J. The effect of cholera toxin on the inhibition of vasopressin-stimulated inositol phospholipid hydrolysis is a cyclic AMP-mediated event at the level of receptor binding. Biochem J. 1989 May 1;259(3):679–684. doi: 10.1042/bj2590679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kolesnick R. N., Paley A. E. 1,2-Diacylglycerols and phorbol esters stimulate phosphatidylcholine metabolism in GH3 pituitary cells. Evidence for separate mechanisms of action. J Biol Chem. 1987 Jul 5;262(19):9204–9210. [PubMed] [Google Scholar]
  11. Liscovitch M., Blusztajn J. K., Freese A., Wurtman R. J. Stimulation of choline release from NG108-15 cells by 12-O-tetradecanoylphorbol 13-acetate. Biochem J. 1987 Jan 1;241(1):81–86. doi: 10.1042/bj2410081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Martin T. W. Formation of diacylglycerol by a phospholipase D-phosphatidate phosphatase pathway specific for phosphatidylcholine in endothelial cells. Biochim Biophys Acta. 1988 Oct 14;962(3):282–296. doi: 10.1016/0005-2760(88)90258-5. [DOI] [PubMed] [Google Scholar]
  13. Moolenaar W. H., Kruijer W., Tilly B. C., Verlaan I., Bierman A. J., de Laat S. W. Growth factor-like action of phosphatidic acid. Nature. 1986 Sep 11;323(6084):171–173. doi: 10.1038/323171a0. [DOI] [PubMed] [Google Scholar]
  14. Muir J. G., Murray A. W. Bombesin and phorbol ester stimulate phosphatidylcholine hydrolysis by phospholipase C: evidence for a role of protein kinase C. J Cell Physiol. 1987 Mar;130(3):382–391. doi: 10.1002/jcp.1041300311. [DOI] [PubMed] [Google Scholar]
  15. Pai J. K., Siegel M. I., Egan R. W., Billah M. M. Activation of phospholipase D by chemotactic peptide in HL-60 granulocytes. Biochem Biophys Res Commun. 1988 Jan 15;150(1):355–364. doi: 10.1016/0006-291x(88)90528-1. [DOI] [PubMed] [Google Scholar]
  16. Palmer S., Wakelam M. J. The Ins(1,4,5)P3 binding site of bovine adrenocortical microsomes: function and regulation. Biochem J. 1989 Jun 1;260(2):593–596. doi: 10.1042/bj2600593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Putney J. W., Jr, Weiss S. J., Van De Walle C. M., Haddas R. A. Is phosphatidic acid a calcium ionophore under neurohumoral control? Nature. 1980 Mar 27;284(5754):345–347. doi: 10.1038/284345a0. [DOI] [PubMed] [Google Scholar]
  18. Rodriguez-Pena A., Rozengurt E. Disappearance of Ca2+-sensitive, phospholipid-dependent protein kinase activity in phorbol ester-treated 3T3 cells. Biochem Biophys Res Commun. 1984 May 16;120(3):1053–1059. doi: 10.1016/s0006-291x(84)80213-2. [DOI] [PubMed] [Google Scholar]
  19. Skipski V. P., Peterson R. F., Barclay M. Quantitative analysis of phospholipids by thin-layer chromatography. Biochem J. 1964 Feb;90(2):374–378. doi: 10.1042/bj0900374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Slivka S. R., Meier K. E., Insel P. A. Alpha 1-adrenergic receptors promote phosphatidylcholine hydrolysis in MDCK-D1 cells. A mechanism for rapid activation of protein kinase C. J Biol Chem. 1988 Sep 5;263(25):12242–12246. [PubMed] [Google Scholar]
  21. Takuwa N., Takuwa Y., Rasmussen H. A tumour promoter, 12-O-tetradecanoylphorbol 13-acetate, increases cellular 1,2-diacylglycerol content through a mechanism other than phosphoinositide hydrolysis in Swiss-mouse 3T3 fibroblasts. Biochem J. 1987 May 1;243(3):647–653. doi: 10.1042/bj2430647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Warden C. H., Friedkin M. Regulation of choline kinase activity and phosphatidylcholine biosynthesis by mitogenic growth factors in 3T3 fibroblasts. J Biol Chem. 1985 May 25;260(10):6006–6011. [PubMed] [Google Scholar]
  23. Warden C. H., Friedkin M. Regulation of phosphatidylcholine biosynthesis by mitogenic growth factors. Biochim Biophys Acta. 1984 Mar 7;792(3):270–280. doi: 10.1016/0005-2760(84)90194-2. [DOI] [PubMed] [Google Scholar]
  24. Welsh C. J., Cao H. T., Chabbott H., Cabot M. C. Vasopressin is the only component of serum-free medium that stimulates phosphatidylcholine hydrolysis and accumulation of diacylglycerol in cultured REF52 cells. Biochem Biophys Res Commun. 1988 Apr 29;152(2):565–572. doi: 10.1016/s0006-291x(88)80075-5. [DOI] [PubMed] [Google Scholar]
  25. Yavin E. Regulation of phospholipid metabolism in differentiating cells from rat brain cerebral hemispheres in culture. Patterns of acetylcholine phosphocholine, and choline phosphoglycerides labeling from (methyl-14C)choline. J Biol Chem. 1976 Mar 10;251(5):1392–1397. [PubMed] [Google Scholar]

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

RESOURCES