Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1992 Jun 1;284(Pt 2):507–512. doi: 10.1042/bj2840507

Oxidized glutathione causes sensitization of calcium release to inositol 1,4,5-trisphosphate in permeabilized hepatocytes.

D C Renard 1, M B Seitz 1, A P Thomas 1
PMCID: PMC1132667  PMID: 1599435

Abstract

The effects of GSSG on Ca2+ mobilization by Ins(1,4,5)P3 were studied in permeabilized rat hepatocytes. Incubation with GSSG (2 mM) increased the sensitivity to Ins(1,4,5)P3 for Ca2+ release, with no effect on the size of the Ca2+ pool that could be released with maximal concentrations of Ins(1,4,5)P3. GSSG decreased the EC50 for Ins(1,4,5)P3 from a control value of 578 +/- 23 nM to 137 +/- 21 nM. GSSG had no effect on the metabolism of Ins(1,4,5)P3 in permeabilized cells, and sensitization of Ca2+ release was still observed when the poorly metabolizable analogue inositol 1,4,5-trisphosphorothioate was used. GSSG did not affect the ATP-dependent Ca2+ pump or the extent of loading of intracellular Ca2+ pools. In addition, the enhancement of Ins(1,4,5)P3-sensitivity by GSSG occurred under conditions where the Ca2+ pumps were blocked with thapsigargin or by chelation of medium Ca2+ just before Ins(1,4,5)P3 addition. The effect of GSSG was time- and dose-dependent, maximal effects being observed after 5 min incubation with 2 mM-GSSG. Cystine mimicked the GSSG-induced increase in Ins(1,4,5)P3-sensitivity, and the effects could be reversed by dithiothreitol (DTT). DTT, GSH glutathione and cysteine had no effect when added alone. Other agents known to react with protein thiols, including N-ethylmaleimide, p-chloromercuribenzoic acid and Ag+, did not affect the sensitivity to Ins(1,4,5)P3, but were inhibitors of ATP-dependent Ca2+ uptake. The data suggest that the sensitivity of the intracellular Ca2+ pools to release by Ins(1,4,5)P3 can be modulated by the formation of mixed disulphides with GSSG or other oxidized thiols.

Full text

PDF
509

Selected References

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

  1. Adunyah S. E., Dean W. L. Effects of sulfhydryl reagents and other inhibitors on Ca2+ transport and inositol trisphosphate-induced Ca2+ release from human platelet membranes. J Biol Chem. 1986 Oct 5;261(28):13071–13075. [PubMed] [Google Scholar]
  2. Bellomo G., Orrenius S. Altered thiol and calcium homeostasis in oxidative hepatocellular injury. Hepatology. 1985 Sep-Oct;5(5):876–882. doi: 10.1002/hep.1840050529. [DOI] [PubMed] [Google Scholar]
  3. Bellomo G., Thor H., Orrenius S. Increase in cytosolic Ca2+ concentration during t-butyl hydroperoxide metabolism by isolated hepatocytes involves NADPH oxidation and mobilization of intracellular Ca2+ stores. FEBS Lett. 1984 Mar 12;168(1):38–42. doi: 10.1016/0014-5793(84)80202-1. [DOI] [PubMed] [Google Scholar]
  4. Burgess G. M., Bird G. S., Obie J. F., Putney J. W., Jr The mechanism for synergism between phospholipase C- and adenylylcyclase-linked hormones in liver. Cyclic AMP-dependent kinase augments inositol trisphosphate-mediated Ca2+ mobilization without increasing the cellular levels of inositol polyphosphates. J Biol Chem. 1991 Mar 15;266(8):4772–4781. [PubMed] [Google Scholar]
  5. Farber J. L., Kyle M. E., Coleman J. B. Mechanisms of cell injury by activated oxygen species. Lab Invest. 1990 Jun;62(6):670–679. [PubMed] [Google Scholar]
  6. Gilbert H. F. Molecular and cellular aspects of thiol-disulfide exchange. Adv Enzymol Relat Areas Mol Biol. 1990;63:69–172. doi: 10.1002/9780470123096.ch2. [DOI] [PubMed] [Google Scholar]
  7. Guillemette G., Segui J. A. Effects of pH, reducing and alkylating reagents on the binding and Ca2+ release activities of inositol 1,4,5-triphosphate in the bovine adrenal cortex. Mol Endocrinol. 1988 Dec;2(12):1249–1255. doi: 10.1210/mend-2-12-1249. [DOI] [PubMed] [Google Scholar]
  8. Jean T., Klee C. B. Calcium modulation of inositol 1,4,5-trisphosphate-induced calcium release from neuroblastoma x glioma hybrid (NG108-15) microsomes. J Biol Chem. 1986 Dec 15;261(35):16414–16420. [PubMed] [Google Scholar]
  9. Joseph S. K., Rice H. L., Williamson J. R. The effect of external calcium and pH on inositol trisphosphate-mediated calcium release from cerebellum microsomal fractions. Biochem J. 1989 Feb 15;258(1):261–265. doi: 10.1042/bj2580261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Joseph S. K., Thomas A. P., Williams R. J., Irvine R. F., Williamson J. R. myo-Inositol 1,4,5-trisphosphate. A second messenger for the hormonal mobilization of intracellular Ca2+ in liver. J Biol Chem. 1984 Mar 10;259(5):3077–3081. [PubMed] [Google Scholar]
  11. Kawanishi T., Blank L. M., Harootunian A. T., Smith M. T., Tsien R. Y. Ca2+ oscillations induced by hormonal stimulation of individual fura-2-loaded hepatocytes. J Biol Chem. 1989 Aug 5;264(22):12859–12866. [PubMed] [Google Scholar]
  12. Masaki N., Kyle M. E., Farber J. L. tert-butyl hydroperoxide kills cultured hepatocytes by peroxidizing membrane lipids. Arch Biochem Biophys. 1989 Mar;269(2):390–399. doi: 10.1016/0003-9861(89)90122-7. [DOI] [PubMed] [Google Scholar]
  13. Mauger J. P., Claret M., Pietri F., Hilly M. Hormonal regulation of inositol 1,4,5-trisphosphate receptor in rat liver. J Biol Chem. 1989 May 25;264(15):8821–8826. [PubMed] [Google Scholar]
  14. Missiaen L., Taylor C. W., Berridge M. J. Spontaneous calcium release from inositol trisphosphate-sensitive calcium stores. Nature. 1991 Jul 18;352(6332):241–244. doi: 10.1038/352241a0. [DOI] [PubMed] [Google Scholar]
  15. Palade P., Dettbarn C., Alderson B., Volpe P. Pharmacologic differentiation between inositol-1,4,5-trisphosphate-induced Ca2+ release and Ca2+- or caffeine-induced Ca2+ release from intracellular membrane systems. Mol Pharmacol. 1989 Oct;36(4):673–680. [PubMed] [Google Scholar]
  16. Pietri F., Hilly M., Mauger J. P. Calcium mediates the interconversion between two states of the liver inositol 1,4,5-trisphosphate receptor. J Biol Chem. 1990 Oct 15;265(29):17478–17485. [PubMed] [Google Scholar]
  17. Prabhu S. D., Salama G. The heavy metal ions Ag+ and Hg2+ trigger calcium release from cardiac sarcoplasmic reticulum. Arch Biochem Biophys. 1990 Feb 15;277(1):47–55. doi: 10.1016/0003-9861(90)90548-d. [DOI] [PubMed] [Google Scholar]
  18. Pruijn F. B., Sibeijn J. P., Bast A. Changes in inositol-1,4,5-trisphosphate binding to hepatic plasma membranes caused by temperature, N-ethylmaleimide and menadione. Biochem Pharmacol. 1990 Nov 1;40(9):1947–1952. doi: 10.1016/0006-2952(90)90223-8. [DOI] [PubMed] [Google Scholar]
  19. Rooney T. A., Renard D. C., Sass E. J., Thomas A. P. Oscillatory cytosolic calcium waves independent of stimulated inositol 1,4,5-trisphosphate formation in hepatocytes. J Biol Chem. 1991 Jul 5;266(19):12272–12282. [PubMed] [Google Scholar]
  20. Rooney T. A., Sass E. J., Thomas A. P. Characterization of cytosolic calcium oscillations induced by phenylephrine and vasopressin in single fura-2-loaded hepatocytes. J Biol Chem. 1989 Oct 15;264(29):17131–17141. [PubMed] [Google Scholar]
  21. Sakaida I., Thomas A. P., Farber J. L. Increases in cytosolic calcium ion concentration can be dissociated from the killing of cultured hepatocytes by tert-butyl hydroperoxide. J Biol Chem. 1991 Jan 15;266(2):717–722. [PubMed] [Google Scholar]
  22. Shan X. Q., Aw T. Y., Jones D. P. Glutathione-dependent protection against oxidative injury. Pharmacol Ther. 1990;47(1):61–71. doi: 10.1016/0163-7258(90)90045-4. [DOI] [PubMed] [Google Scholar]
  23. Supattapone S., Worley P. F., Baraban J. M., Snyder S. H. Solubilization, purification, and characterization of an inositol trisphosphate receptor. J Biol Chem. 1988 Jan 25;263(3):1530–1534. [PubMed] [Google Scholar]
  24. Thastrup O., Cullen P. J., Drøbak B. K., Hanley M. R., Dawson A. P. Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2(+)-ATPase. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2466–2470. doi: 10.1073/pnas.87.7.2466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Thomas A. P. Enhancement of the inositol 1,4,5-trisphosphate-releasable Ca2+ pool by GTP in permeabilized hepatocytes. J Biol Chem. 1988 Feb 25;263(6):2704–2711. [PubMed] [Google Scholar]
  26. Woods N. M., Cuthbertson K. S., Cobbold P. H. Repetitive transient rises in cytoplasmic free calcium in hormone-stimulated hepatocytes. Nature. 1986 Feb 13;319(6054):600–602. doi: 10.1038/319600a0. [DOI] [PubMed] [Google Scholar]
  27. Yang C. M., Lee H. C. Effects of sulfhydryl reagents on [3H] inositol trisphosphate binding to dog cerebellar membranes. J Recept Res. 1989;9(2):159–169. doi: 10.3109/10799898909066051. [DOI] [PubMed] [Google Scholar]

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

RESOURCES