Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1975 Nov;12(5):964–968. doi: 10.1128/iai.12.5.964-968.1975

Selective inhibition of cholera toxin- and catecholamine-stimulated lipolysis by blocking agents.

M S Katz, W B Greenough 3rd
PMCID: PMC415383  PMID: 1193734

Abstract

Vibrio cholerae enterotoxin stimulates lipolysis in rat epididymal fat cell suspensions. Like hormones this toxin increases adenylate cyclase activity, raising levels of cyclic adenosine 3',5'-monophosphate (cAMP), which activates a cellular lipase. Using specific blocking agents, we studied the responses to the adrenergic lipolytic hormones epinephrine, norepinephrine, and isoproterenol, and to cholera toxin. All stimulators were used at 100 x threshold dose. Propranolol (34 muM), a beta blocking agent, inhibited epinephrine stimulation (P less than 0.001) but not that of toxin (P greater than 0.2). Choleragenoid (25 mug/ml), a natural toxoid of cholera toxin, blocked stimulation by toxin (P less than 0.001) but not that of the adrenergic agents (P greater than 0.2). A beta blocker, practolol (3 mM), inhibited stimulation by the catecholamines tested (P less than 0.005) but not that of toxin (P greater than 0.05). Higher concentrations of propranolol (340 muM) and the alpha blocking agents phenoxybenzamine (3 mM) and phentolamine (1.6 mM) inhibited all agonists (P less than 0.001). The response to theophylline was inhibited by all blockers (P less than 0.05) except propranolol at the lower concentration (34 muM). A combined beta and alpha blockade using propranolol and epinephrine together did not inhibit toxin-mediated lipolysis. It appears that stimulation by cholera toxin is independent of beta adrenergic receptors. A major inhibition of theophylline-mediated lipolysis by alpha blocking drugs indicated a nonspecific effect of these agents at the concentrations used. The uninhibited response to toxin in the presence of propranolol and epinephrine suggests a lack of relationship of the toxin receptor to either alpha or beta receptors.

Full text

PDF
964

Selected References

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

  1. Beavo J. A., Rogers N. L., Crofford O. B., Hardman J. G., Sutherland E. W., Newman E. V. Effects of xanthine derivatives on lipolysis and on adenosine 3',5'-monophosphate phosphodiesterase activity. Mol Pharmacol. 1970 Nov;6(6):597–603. [PubMed] [Google Scholar]
  2. Beckman B., Flores J., Witkum P. A., Sharp G. W. Studies on the mode of action of cholera toxin. Effects on solubilized adenylate cyclase. J Clin Invest. 1974 Apr;53(4):1202–1205. doi: 10.1172/JCI107660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burns T. W., Langley P. E., Robison G. A. Adrenergic receptors and cyclic AMP in the regulation of human adipose tissue lipolysis. Ann N Y Acad Sci. 1971 Dec 30;185:115–128. doi: 10.1111/j.1749-6632.1971.tb45242.x. [DOI] [PubMed] [Google Scholar]
  4. Cuatrecasas P. Cholera toxin-fat cell interaction and the mechanism of activation of the lipolytic response. Biochemistry. 1973 Aug 28;12(18):3567–3577. doi: 10.1021/bi00742a033. [DOI] [PubMed] [Google Scholar]
  5. Cuatrecasas P. Gangliosides and membrane receptors for cholera toxin. Biochemistry. 1973 Aug 28;12(18):3558–3566. doi: 10.1021/bi00742a032. [DOI] [PubMed] [Google Scholar]
  6. Cuatrecasas P., Parikh I., Hollenberg M. D. Affinity chromatography and structural analysis of Vibrio cholerae enterotoxin-ganglioside agarose and the biological effects of ganglioside-containing soluble polymers. Biochemistry. 1973 Oct 9;12(21):4253–4264. doi: 10.1021/bi00745a033. [DOI] [PubMed] [Google Scholar]
  7. Cuatrecasas P. Vibrio cholerae choleragenoid. Mechanism of inhibition of cholera toxin action. Biochemistry. 1973 Aug 28;12(18):3577–3581. doi: 10.1021/bi00742a034. [DOI] [PubMed] [Google Scholar]
  8. Curlin C. T., Mosley W. H., Greenough W. B., 3rd Cholera antitoxin titrations: a comparative study of fat-cell, ileal-loop, and rabbit-skin assays. J Infect Dis. 1973 Mar;127(3):294–298. doi: 10.1093/infdis/127.3.294. [DOI] [PubMed] [Google Scholar]
  9. Field M. Mode of action of cholera toxin: stabilization of catecholamine-sensitive adenylate cyclase in turkey erythrocytes. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3299–3303. doi: 10.1073/pnas.71.8.3299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hewlett E. L., Guerrant R. L., Evans D. J., Jr, Greenough W. B., 3rd Toxins of Vibrio cholerae and Escherichia coli stimulate adenyl cyclase in rat fat cells. Nature. 1974 May 24;249(455):371–373. doi: 10.1038/249371a0. [DOI] [PubMed] [Google Scholar]
  11. Heyningen S Van Cholera toxin: interaction of subunits with ganglioside GM1. Science. 1974 Feb 15;183(4125):656–657. doi: 10.1126/science.183.4125.656. [DOI] [PubMed] [Google Scholar]
  12. Holmgren J., Lönnroth I., Svennerholm L. Tissue receptor for cholera exotoxin: postulated structure from studies with GM1 ganglioside and related glycolipids. Infect Immun. 1973 Aug;8(2):208–214. doi: 10.1128/iai.8.2.208-214.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Khoo J. C., Aquino A. A., Steinberg D. The mechanism of activation of hormone-sensitive lipase in human adipose tissue. J Clin Invest. 1974 Apr;53(4):1124–1131. doi: 10.1172/JCI107650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. King C. A., Van Heyningen W. E. Deactivation of cholera toxin by a sialidase-resistant monosialosylganglioside. J Infect Dis. 1973 Jun;127(6):639–647. doi: 10.1093/infdis/127.6.639. [DOI] [PubMed] [Google Scholar]
  15. Pierce N. F. Differential inhibitory effects of cholera toxoids and ganglioside on the enterotoxins of Vibrio cholerae and Escherichia coli. J Exp Med. 1973 Apr 1;137(4):1009–1023. doi: 10.1084/jem.137.4.1009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pierce N. F., Greenough W. B., 3rd, Carpenter C. C., Jr Vibrio cholerae enterotoxin and its mode of action. Bacteriol Rev. 1971 Mar;35(1):1–13. doi: 10.1128/br.35.1.1-13.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. RODBELL M. METABOLISM OF ISOLATED FAT CELLS. I. EFFECTS OF HORMONES ON GLUCOSE METABOLISM AND LIPOLYSIS. J Biol Chem. 1964 Feb;239:375–380. [PubMed] [Google Scholar]
  18. Sheppard H. Inhibition of norepinephrine stimulated adenyl cyclase by theophylline. Nature. 1970 Nov 7;228(5271):567–568. doi: 10.1038/228567a0. [DOI] [PubMed] [Google Scholar]
  19. Vaughan M., Pierce N. F., Greenough W. B., 3rd Stimulation of glycerol production in fat cells by cholera toxin. Nature. 1970 May 16;226(5246):658–659. doi: 10.1038/226658a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES