Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1977 Sep 1;165(3):447–454. doi: 10.1042/bj1650447

The effects of amphiphilic cationic drugs and inorganic cations on the activity of phosphatidate phosphohydrolase.

M Bowley, J Cooling, S L Burditt, D N Brindley
PMCID: PMC1164926  PMID: 200224

Abstract

1. Phosphatidate phosphohydrolase from the particle-free supernatant of rat liver was assayed by using emulsions of phosphatidate as substrate. 2. The inhibition of the phosphohydrolase by chlorpromazine was of a competitive type with respect to phosphatidate. The potency of various amphiphilic cationic drugs as inhibitors of this reaction was related to their partition coefficients into a phosphatidate emulsion. 3. The effect of chlorpromazine on the phosphohydrolase activity was complementary rather than antagonistic towards Mg2+. Chlorpromazine stimulated the phosphohydrolase activity in the absence of added Mg2+ and was able to replace the requirement for Mg2+. However, at optimum concentrations of Mg2+, chlorpromazine inhibited the reaction, as did Ca2+. The phosphohydrolase activity was also stimulated by Co2+ and to a lesser extent by Mn2+, Fe2+, Fe3+, Ca2+, spermine and spermidine when Mg2+ was not added to the assays. 4. It is concluded that the inhibition of phosphatidate phosphohydrolase by amphiphilic cations can largely be explained by the interaction of these compounds with phosphatidate, which changes the physical properties of the lipid, making it less available for conversion into diacylglycerol. 5. The implications of these results to the effects of amphiphilic cations in redirecting glycerolipid synthesis at the level of phosphatidate are discussed.

Full text

PDF
447

Selected References

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

  1. Abdel-Latif A. A., Smith J. P. Effects of DL-propranolol on the synthesis of glycerolipids by rabbit iris muscle. Biochem Pharmacol. 1976 Aug 1;25(15):1697–1704. doi: 10.1016/0006-2952(76)90401-9. [DOI] [PubMed] [Google Scholar]
  2. Allan D., Michell R. H. Enhanced synthesis de novo of phosphatidylinositol in lymphocytes treated with cationic amphiphilic drugs. Biochem J. 1975 Jun;148(3):471–478. doi: 10.1042/bj1480471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  4. Bangham A. D., Standish M. M., Miller N. Cation permeability of phospholipid model membranes: effect of narcotics. Nature. 1965 Dec 25;208(5017):1295–1297. doi: 10.1038/2081295a0. [DOI] [PubMed] [Google Scholar]
  5. Bazán N. G., De Boschero M. G., Giusto N. M., De Bazán H. E. De novo glycerolipid biosynthesis in the toad and cattle retina. Redirecting of the pathway by propranolol and phentolamine. Adv Exp Med Biol. 1976;72:139–148. doi: 10.1007/978-1-4684-0955-0_13. [DOI] [PubMed] [Google Scholar]
  6. Bickel M. H., Steele J. W. Binding of basic and acidic drugs to rat tissue subcellular fractions. Chem Biol Interact. 1974 Mar;8(3):151–162. doi: 10.1016/0009-2797(74)90037-4. [DOI] [PubMed] [Google Scholar]
  7. Blank M. L., Wykle R. L., Alper S., Snyder F. Microsomal synthesis of the ether analogs of triacylglycerols. Acyl CoA:alkylacylglycerol and acyl CoA:alk-i-enylacylglycerol acyltransferases in tumors and liver. Biochim Biophys Acta. 1974 Jun 26;348(3):397–403. doi: 10.1016/0005-2760(74)90219-7. [DOI] [PubMed] [Google Scholar]
  8. Brindley D. N., Allan D., Michell R. H. Letter: The redirection of glyceride and phospholipid synthesis by drugs including chlorpromazine, fenfluramine, imipramine, mepyramine and local anaesthetics. J Pharm Pharmacol. 1975 Jun;27(6):462–464. [PubMed] [Google Scholar]
  9. Brindley D. N., Bowley M., Burditt S., Pritchard H., Lloyd-Davies K. A., Boucrot P. The effects of administering N-(2-benzoyloxyethyl) norfenfluramine to rats on the hepatic synthesis of glycerolipids. J Pharm Pharmacol. 1976 Sep;28(9):676–682. doi: 10.1111/j.2042-7158.1976.tb02835.x. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Brindley D. N. The intracellular phase of fat absorption. Biomembranes. 1974;4B(0):621–671. doi: 10.1007/978-1-4684-3336-4_2. [DOI] [PubMed] [Google Scholar]
  12. COLEMAN R., HUEBSCHER G. Metabolism of phospholipids. V. Studies of phosphatidic acid phosphatase. Biochim Biophys Acta. 1962 Jan 29;56:479–490. doi: 10.1016/0006-3002(62)90600-5. [DOI] [PubMed] [Google Scholar]
  13. Call F. L., 2nd, Williams W. J. Phosphatidate phosphatase in human platelets. J Lab Clin Med. 1973 Oct;82(4):663–673. [PubMed] [Google Scholar]
  14. Caras I., Shapiro B. Partial purification and properties of microsomal phosphatidate phosphohydrolase from rat liver. Biochim Biophys Acta. 1975 Nov 21;409(2):201–211. doi: 10.1016/0005-2760(75)90154-x. [DOI] [PubMed] [Google Scholar]
  15. Carey M. C., Hirom P. C., Small D. M. A study of the physicochemical interactions between biliary lipids and chlorpromazine hydrochloride. Bile-salt precipitation as a mechanism of phenothiazine-induced bile secretory failure. Biochem J. 1976 Mar 1;153(3):519–531. doi: 10.1042/bj1530519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dodds P. F., Gurr M. I., Brindley D. N. The glycerol phosphate, dihydroxyacetone phosphate and monoacylglycerol pathways of glycerolipid synthesis in rat adipose-tissue homogenates. Biochem J. 1976 Dec 15;160(3):693–700. doi: 10.1042/bj1600693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Eichberg J., Hauser G. Stimulation by local anesthetics of the metabolism of acidic phospholipids in the rat pineal gland. Biochem Biophys Res Commun. 1974 Oct 23;60(4):1460–1467. doi: 10.1016/0006-291x(74)90362-3. [DOI] [PubMed] [Google Scholar]
  18. Fallon H. J., Lamb R. G., Jamdar S. C. Phosphatidate phosphohydrolase and the regulation of glycerolipid biosynthesis. Biochem Soc Trans. 1977;5(1):37–40. doi: 10.1042/bst0050037. [DOI] [PubMed] [Google Scholar]
  19. Fernández M. S., Cerbón J. The importance of the hydrophobic interactions of local anesthetics in the displacement of polyvalent cations from artificial lipid membranes. Biochim Biophys Acta. 1973 Feb 27;298(1):8–14. doi: 10.1016/0005-2736(73)90003-5. [DOI] [PubMed] [Google Scholar]
  20. Giotta G. J., Chan D. S., Wang H. H. Binding of spin-labeled local anesthetics to phosphatidylcholine and phosphatidylserine liposomes. Arch Biochem Biophys. 1974 Aug;163(2):453–458. doi: 10.1016/0003-9861(74)90501-3. [DOI] [PubMed] [Google Scholar]
  21. Hajra A. K., Seguin E. B., Agranoff B. W. Rapid labeling of mitochondrial lipids by labeled orthophosphate and adenosine triphosphate. J Biol Chem. 1968 Apr 10;243(7):1609–1616. [PubMed] [Google Scholar]
  22. Hansch C., Glave W. R. Structure-activity relationships in membrane-perturbing agents. Hemolytic, narcotic, and antibacterial compounds. Mol Pharmacol. 1971 May;7(3):337–354. [PubMed] [Google Scholar]
  23. Hauser H., Penkett S. A., Chapman D. Nuclear magnetic resonance spectroscopic studies of procaine hydrochloride and tetracaine hydrochloride at lipid-water interfaces. Biochim Biophys Acta. 1969;183(3):466–475. doi: 10.1016/0005-2736(69)90161-8. [DOI] [PubMed] [Google Scholar]
  24. Hosaka K., Yamashita S., Numa S. Partial purification, properties, and subcellulsr distribution of rat liver phosphatidate phosphatase. J Biochem. 1975 Mar;77(3):501–509. doi: 10.1093/oxfordjournals.jbchem.a130751. [DOI] [PubMed] [Google Scholar]
  25. Hübscher G., Brindley D. N., Smith M. E., Sedgwick B. Stimulation of biosynthesis of glyceride. Nature. 1967 Nov 4;216(5114):449–453. doi: 10.1038/216449a0. [DOI] [PubMed] [Google Scholar]
  26. Hübscher G., West G. R., Brindley D. N. Studies on the fractionation of mucosal homogenates from the small intestine. Biochem J. 1965 Dec;97(3):629–642. doi: 10.1042/bj0970629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Jamdar S. C., Fallon H. J. Glycerolipid synthesis in rat adipose tissue. II. Properties and distribution of phosphatidate phosphatase. J Lipid Res. 1973 Sep;14(5):517–524. [PubMed] [Google Scholar]
  28. Kako K. J., Patterson S. D. Phosphatidate phosphohydrolase and palmitoyl-coenzyme A hydrolase in cardiac subcellular fractions of hyperthyroid rabbits and cardiomyopathic hamsters. Biochem J. 1975 Nov;152(2):313–323. doi: 10.1042/bj1520313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kent C., Vagelos P. R. Phosphatidic acid phosphatase and phospholipdase A activities in plasma membranes from fusing muscle cells. Biochim Biophys Acta. 1976 Jun 17;436(2):377–386. doi: 10.1016/0005-2736(76)90201-7. [DOI] [PubMed] [Google Scholar]
  30. Kwant W. O., Seeman P. The membrane concentration of a local anesthetic (chlorpromazine). Biochim Biophys Acta. 1969;183(3):530–543. doi: 10.1016/0005-2736(69)90167-9. [DOI] [PubMed] [Google Scholar]
  31. Lee A. G. Interactions between anaesthetics and lipid mixtures. Amines. Biochim Biophys Acta. 1976 Sep 21;448(1):34–44. doi: 10.1016/0005-2736(76)90074-2. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Michell R. H., Allan D., Bowley M., Brindley D. N. A possible metabolic explanation for drug-induced phospholipidosis. J Pharm Pharmacol. 1976 Apr;28(4):331–332. doi: 10.1111/j.2042-7158.1976.tb04172.x. [DOI] [PubMed] [Google Scholar]
  34. Mitchell M. P., Brindley D. N., Hübscher G. Properties of phosphatidate phosphohydrolase. Eur J Biochem. 1971 Jan;18(2):214–220. doi: 10.1111/j.1432-1033.1971.tb01233.x. [DOI] [PubMed] [Google Scholar]
  35. Papahadjopoulos D., Jacobson K., Poste G., Shepherd G. Effects of local anesthetics on membrane properties. I. Changes in the fluidity of phospholipid bilayers. Biochim Biophys Acta. 1975 Jul 18;394(4):504–519. doi: 10.1016/0005-2736(75)90137-6. [DOI] [PubMed] [Google Scholar]
  36. Papahadjopoulos D. Phospholipid model membranes. 3. Antagonistic effects of Ca2+ and local anesthetics on the permeability of phosphatidylserine vesicles. Biochim Biophys Acta. 1970 Sep 15;211(3):467–477. doi: 10.1016/0005-2736(70)90252-x. [DOI] [PubMed] [Google Scholar]
  37. Papahadjopoulos D. Studies on the mechanism of action of local anesthetics with phospholipid model membranes. Biochim Biophys Acta. 1972 Apr 18;265(2):169–186. doi: 10.1016/0304-4157(72)90001-9. [DOI] [PubMed] [Google Scholar]
  38. Pritchard P. H., Brindley D. N. Studies on the ethanol-induced changes in glycerolipid synthesis in rats and their partial reversal by N-(2-benzoyloxyethyl)norfenfluramine (benfluorex). J Pharm Pharmacol. 1977 Jun;29(6):343–349. doi: 10.1111/j.2042-7158.1977.tb11332.x. [DOI] [PubMed] [Google Scholar]
  39. RAAFLAUB J. Applications of metal buffers and metal indicators in biochemistry. Methods Biochem Anal. 1956;3:301–325. doi: 10.1002/9780470110195.ch10. [DOI] [PubMed] [Google Scholar]
  40. Ragazzi M., Gorio A., Peluchetti D. Influence of calcium on the interaction of anesthetic drugs with artificial phospholipid membranes. Experientia. 1975 May 15;31(5):567–568. doi: 10.1007/BF01932462. [DOI] [PubMed] [Google Scholar]
  41. Scherphof G., Westenberg H. Stimulation and inhibition of pancreatic phospholipase A2 by local anesthetics as a result of their interaction with the substrate. Biochim Biophys Acta. 1975 Sep 19;398(3):442–451. doi: 10.1016/0005-2760(75)90195-2. [DOI] [PubMed] [Google Scholar]
  42. Sedgwick B., Hübscher G. Metabolism of phospholipids. IX. Phosphatidate phosphohydrolase in rat liver. Biochim Biophys Acta. 1965 Jul 7;106(1):63–77. doi: 10.1016/0005-2760(65)90096-2. [DOI] [PubMed] [Google Scholar]
  43. Sedgwick B., Hübscher G. Metabolism of phospholipids. X. Partial purification and properties of a soluble phosphatidate phosphohydrolase from rat liver. Biochim Biophys Acta. 1967 Oct 2;144(2):397–408. [PubMed] [Google Scholar]
  44. Seydel J. K., Wassermann O. NMR-studies on the molecular basis of drug-induced phospholipidosis--II. Interaction between several amphiphilic drugs and phospholipids. Biochem Pharmacol. 1976 Nov 1;25(21):2357–2364. doi: 10.1016/0006-2952(76)90028-9. [DOI] [PubMed] [Google Scholar]
  45. Sluyterman L. A., Wijdenes J. An unusual type of enzyme inhibition. Biochim Biophys Acta. 1973 Oct 10;321(2):697–699. doi: 10.1016/0005-2744(73)90218-0. [DOI] [PubMed] [Google Scholar]
  46. Smith M. E., Sedgwick B., Brindley D. N., Hübscer G. The role of phosphatidate phosphohydrolase in glyceride biosynthesis. Eur J Biochem. 1967 Dec;3(1):70–77. doi: 10.1111/j.1432-1033.1967.tb19499.x. [DOI] [PubMed] [Google Scholar]
  47. Sturton R. G., Brindley D. N. Factors controlling the activities of phosphatidate phosphohydrolase and phosphatidate cytidylyltransferase. The effects of chlorpromazine, demethylimipramine, cinchocaine, norfenfluramine, mepyramine and magnesium ions. Biochem J. 1977 Jan 15;162(1):25–32. doi: 10.1042/bj1620025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. 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]
  49. Traynor J. R., Kunze H. Effects of local anaesthetics on the lipase of Rhizopus arrhizus. Biochim Biophys Acta. 1976 Feb 23;424(2):246–252. doi: 10.1016/0005-2760(76)90192-2. [DOI] [PubMed] [Google Scholar]
  50. Ueda I., Wada T. Determination of inorganic phosphate by the molybdovanadate method in the presence of ATP and some interfering organic bases. Anal Biochem. 1970 Sep;37(1):169–174. doi: 10.1016/0003-2697(70)90273-3. [DOI] [PubMed] [Google Scholar]
  51. Vavrecka M., Mitchell M. P., Hübscher G. The effect of starvation on the incorporation of palmitate into glycerides and phospholipids of rat liver homogenates. Biochem J. 1969 Nov;115(2):139–145. doi: 10.1042/bj1150139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. WILKINSON G. N. Statistical estimations in enzyme kinetics. Biochem J. 1961 Aug;80:324–332. doi: 10.1042/bj0800324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wilschut J. C., Regts J., Westenberg H., Scherphof G. Hydrolysis of phosphatidylcholine liposomes by phospholipases A2. Effects of the local anesthetic dibucaine. Biochim Biophys Acta. 1976 Apr 16;433(1):20–31. doi: 10.1016/0005-2736(76)90174-7. [DOI] [PubMed] [Google Scholar]

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

RESOURCES