Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1994 Feb;62(2):717–721. doi: 10.1128/iai.62.2.717-721.1994

Regulation of Clostridium perfringens alpha-toxin-activated phospholipase C in rabbit erythrocyte membranes.

J Sakurai 1, S Ochi 1, H Tanaka 1
PMCID: PMC186165  PMID: 8300231

Abstract

The rapid phosphatidic acid (PA) formation induced by Clostridium perfringens alpha-toxin was stimulated by AlF4- in rabbit erythrocyte membranes. GTP[gamma S] [guanosine 5'-O-(3-thiotriphosphate)] stimulated the rapid 1,2-diacylglycerol formation and inositol 1,4,5-trisphosphate release induced by the toxin. On the other hand, treatment of erythrocyte lysates with phorbol 12-myristate 13-acetate (PMA) resulted in inhibition of toxin-induced PA production, and long-term PMA or 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) treatment of the lysates led to stimulation of PA formation. Furthermore, treatment of erythrocytes with the toxin caused an increase of protein kinase C activity in membrane fractions. The results suggest that toxin-induced PA formation is mediated by endogenous phospholipase C regulated through GTP-binding protein and protein kinase C in rabbit erythrocytes.

Full text

PDF

Selected References

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

  1. Agwu D. E., McPhail L. C., Chabot M. C., Daniel L. W., Wykle R. L., McCall C. E. Choline-linked phosphoglycerides. A source of phosphatidic acid and diglycerides in stimulated neutrophils. J Biol Chem. 1989 Jan 25;264(3):1405–1413. [PubMed] [Google Scholar]
  2. Bigay J., Deterre P., Pfister C., Chabre M. Fluoride complexes of aluminium or beryllium act on G-proteins as reversibly bound analogues of the gamma phosphate of GTP. EMBO J. 1987 Oct;6(10):2907–2913. doi: 10.1002/j.1460-2075.1987.tb02594.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brass L. F., Laposata M., Banga H. S., Rittenhouse S. E. Regulation of the phosphoinositide hydrolysis pathway in thrombin-stimulated platelets by a pertussis toxin-sensitive guanine nucleotide-binding protein. Evaluation of its contribution to platelet activation and comparisons with the adenylate cyclase inhibitory protein, Gi. J Biol Chem. 1986 Dec 25;261(36):16838–16847. [PubMed] [Google Scholar]
  4. 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]
  5. Fujii Y., Nomura S., Oshita Y., Sakurai J. Excitatory effect of Clostridium perfringens alpha toxin on the rat isolated aorta. Br J Pharmacol. 1986 Jul;88(3):531–539. doi: 10.1111/j.1476-5381.1986.tb10233.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fujii Y., Sakurai J. Contraction of the rat isolated aorta caused by Clostridium perfringens alpha toxin (phospholipase C): evidence for the involvement of arachidonic acid metabolism. Br J Pharmacol. 1989 May;97(1):119–124. doi: 10.1111/j.1476-5381.1989.tb11931.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gonzalez-Sastre F., Folch-Pi J. Thin-layer chromatography of the phosphoinositides. J Lipid Res. 1968 Jul;9(4):532–533. [PubMed] [Google Scholar]
  8. Hostetler K. Y., Gardner M. F., Aldern K. A. Assay of phospholipases C and D in presence of other lipid hydrolases. Methods Enzymol. 1991;197:125–134. doi: 10.1016/0076-6879(91)97139-p. [DOI] [PubMed] [Google Scholar]
  9. Kikkawa U., Takai Y., Tanaka Y., Miyake R., Nishizuka Y. Protein kinase C as a possible receptor protein of tumor-promoting phorbol esters. J Biol Chem. 1983 Oct 10;258(19):11442–11445. [PubMed] [Google Scholar]
  10. Kopf G. S., Woolkalis M. J. ADP-ribosylation of G proteins with pertussis toxin. Methods Enzymol. 1991;195:257–266. doi: 10.1016/0076-6879(91)95171-f. [DOI] [PubMed] [Google Scholar]
  11. Krug E. L., Kent C. Phospholipase C from Clostridium perfringens: preparation and characterization of homogeneous enzyme. Arch Biochem Biophys. 1984 Jun;231(2):400–410. doi: 10.1016/0003-9861(84)90403-x. [DOI] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Lawrence W. D., Schoenl M., Davis P. J. Stimulation in vitro of rabbit erythrocyte cytosol phospholipid-dependent protein kinase activity. A novel action of thyroid hormone. J Biol Chem. 1989 Mar 25;264(9):4766–4768. [PubMed] [Google Scholar]
  15. Leeb-Lundberg L. M., Cotecchia S., Lomasney J. W., DeBernardis J. F., Lefkowitz R. J., Caron M. G. Phorbol esters promote alpha 1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5651–5655. doi: 10.1073/pnas.82.17.5651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. MACFARLANE M. G. The biochemistry of bacterial toxins; variation in haemolytic activity of immunologically distinct lecithinases towards erythrocytes from different species. Biochem J. 1950 Sep;47(3):270–279. doi: 10.1042/bj0470270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. MacIntyre D. E., McNicol A., Drummond A. H. Tumour-promoting phorbol esters inhibit agonist-induced phosphatidate formation and Ca2+ flux in human platelets. FEBS Lett. 1985 Jan 28;180(2):160–164. doi: 10.1016/0014-5793(85)81063-2. [DOI] [PubMed] [Google Scholar]
  18. Murphy A. C., Rozengurt E. Pasteurella multocida toxin selectively facilitates phosphatidylinositol 4,5-bisphosphate hydrolysis by bombesin, vasopressin, and endothelin. Requirement for a functional G protein. J Biol Chem. 1992 Dec 15;267(35):25296–25303. [PubMed] [Google Scholar]
  19. Nakamura T., Ui M. Simultaneous inhibitions of inositol phospholipid breakdown, arachidonic acid release, and histamine secretion in mast cells by islet-activating protein, pertussis toxin. A possible involvement of the toxin-specific substrate in the Ca2+-mobilizing receptor-mediated biosignaling system. J Biol Chem. 1985 Mar 25;260(6):3584–3593. [PubMed] [Google Scholar]
  20. Nishizuka Y. Turnover of inositol phospholipids and signal transduction. Science. 1984 Sep 21;225(4668):1365–1370. doi: 10.1126/science.6147898. [DOI] [PubMed] [Google Scholar]
  21. Orellana S. A., Solski P. A., Brown J. H. Phorbol ester inhibits phosphoinositide hydrolysis and calcium mobilization in cultured astrocytoma cells. J Biol Chem. 1985 May 10;260(9):5236–5239. [PubMed] [Google Scholar]
  22. Osugi T., Imaizumi T., Mizushima A., Uchida S., Yoshida H. Phorbol ester inhibits bradykinin-stimulated inositol trisphosphate formation and calcium mobilization in neuroblastoma x glioma hybrid NG108-15 cells. J Pharmacol Exp Ther. 1987 Feb;240(2):617–622. [PubMed] [Google Scholar]
  23. Palmer S., Hughes K. T., Lee D. Y., Wakelam M. J. Development of a novel, Ins(1,4,5)P3-specific binding assay. Its use to determine the intracellular concentration of Ins(1,4,5)P3 in unstimulated and vasopressin-stimulated rat hepatocytes. Cell Signal. 1989;1(2):147–156. doi: 10.1016/0898-6568(89)90004-1. [DOI] [PubMed] [Google Scholar]
  24. Paris S., Pouysségur J. Further evidence for a phospholipase C-coupled G protein in hamster fibroblasts. Induction of inositol phosphate formation by fluoroaluminate and vanadate and inhibition by pertussis toxin. J Biol Chem. 1987 Feb 15;262(5):1970–1976. [PubMed] [Google Scholar]
  25. Preiss J., Loomis C. R., Bishop W. R., Stein R., Niedel J. E., Bell R. M. Quantitative measurement of sn-1,2-diacylglycerols present in platelets, hepatocytes, and ras- and sis-transformed normal rat kidney cells. J Biol Chem. 1986 Jul 5;261(19):8597–8600. [PubMed] [Google Scholar]
  26. Ryu S. H., Kim U. H., Wahl M. I., Brown A. B., Carpenter G., Huang K. P., Rhee S. G. Feedback regulation of phospholipase C-beta by protein kinase C. J Biol Chem. 1990 Oct 15;265(29):17941–17945. [PubMed] [Google Scholar]
  27. Sabban E., Laster Y., Loyter A. Resolution of the hemolytic and the hydrolytic activities of phospholipase-C preparation from Clostridium perfringens. Eur J Biochem. 1972 Jul 24;28(3):373–380. doi: 10.1111/j.1432-1033.1972.tb01923.x. [DOI] [PubMed] [Google Scholar]
  28. Sakurai J., Fujii Y., Shirotani M. Contraction induced by Clostridium perfringens alpha toxin in the isolated rat ileum. Toxicon. 1990;28(4):411–418. doi: 10.1016/0041-0101(90)90079-m. [DOI] [PubMed] [Google Scholar]
  29. Sakurai J., Fujii Y., Torii K., Kobayashi K. Dissociation of various biological activities of Clostridium perfringens alpha toxin by chemical modification. Toxicon. 1989;27(3):317–323. doi: 10.1016/0041-0101(89)90179-7. [DOI] [PubMed] [Google Scholar]
  30. Sakurai J., Ochi S., Tanaka H. Evidence for coupling of Clostridium perfringens alpha-toxin-induced hemolysis to stimulated phosphatidic acid formation in rabbit erythrocytes. Infect Immun. 1993 Sep;61(9):3711–3718. doi: 10.1128/iai.61.9.3711-3718.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sato H., Chiba J., Sato Y. Monoclonal antibodies against alpha toxin of Clostridium perfringens. FEMS Microbiol Lett. 1989 May;50(1-2):173–176. doi: 10.1016/0378-1097(89)90480-1. [DOI] [PubMed] [Google Scholar]
  32. Talwar H. S., Fisher G. J., Voorhees J. J. Bradykinin induces phosphoinositide turnover, 1,2-diglyceride formation, and growth in cultured adult human keratinocytes. J Invest Dermatol. 1990 Dec;95(6):705–710. doi: 10.1111/1523-1747.ep12514507. [DOI] [PubMed] [Google Scholar]
  33. Van Heyningen W. E. The biochemistry of the gas gangrene toxins: Partial purification of the toxins of Cl. welchii, type A. Separation of alpha and theta toxins. Biochem J. 1941 Nov;35(10-11):1257–1269. doi: 10.1042/bj0351257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Volpi M., Naccache P. H., Molski T. F., Shefcyk J., Huang C. K., Marsh M. L., Munoz J., Becker E. L., Sha'afi R. I. Pertussis toxin inhibits fMet-Leu-Phe- but not phorbol ester-stimulated changes in rabbit neutrophils: role of G proteins in excitation response coupling. Proc Natl Acad Sci U S A. 1985 May;82(9):2708–2712. doi: 10.1073/pnas.82.9.2708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wada S., Yasutomo Y., Kosano H., Kugai N., Nagata N. The effect of PGF2 alpha on parathyroid hormone-stimulated cyclic AMP production in mouse osteoblastic cell, MC3T3E1. Biochim Biophys Acta. 1991 May 24;1074(1):182–188. doi: 10.1016/0304-4165(91)90059-p. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES