Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1988 Nov 15;256(1):117–124. doi: 10.1042/bj2560117

Characterization of Ca2+ fluxes in rat liver plasma-membrane vesicles.

C Dargemont 1, M Hilly 1, M Claret 1, J P Mauger 1
PMCID: PMC1135376  PMID: 3265619

Abstract

Inside-out plasma-membrane vesicles isolated from rat liver [Prpic, Green, Blackmore & Exton (1984) J. Biol. Chem. 259, 1382-1385] accumulated a substantial amount of 45Ca2+ when they were incubated in a medium whose ionic composition and pH mimicked those of cytosol and which contained MgATP. The Vmax of the initial 45Ca2+ uptake rate was 2.9 +/- 0.6 nmol/min per mg and the Km for Ca2+ was 0.50 +/- 0.08 microM. The ATP-dependent 45Ca2+ uptake by inside-out plasma-membrane vesicles was about 20 times more sensitive to saponin than was the ATP-dependent uptake by a microsomal preparation. The 45Ca2+ efflux from the inside-out vesicles, which is equivalent to the Ca2+ influx in intact cells, was increased when the free Ca2+ concentration in the medium was decreased. The Ca2+ antagonists La3+ and Co2+ inhibited the 45Ca2+ efflux from the vesicles. Neomycin stimulated the Ca2+ efflux in the presence of either a high or a low free Ca2+ concentration. These results confirm that polyvalent cations regulate Ca2+ fluxes through the plasma membrane.

Full text

PDF
118

Selected References

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

  1. Altin J. G., Bygrave F. L. The influx of Ca2+ induced by the administration of glucagon and Ca2+-mobilizing agents to the perfused rat liver could involve at least two separate pathways. Biochem J. 1987 Feb 15;242(1):43–50. doi: 10.1042/bj2420043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berridge M. J., Irvine R. F. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature. 1984 Nov 22;312(5992):315–321. doi: 10.1038/312315a0. [DOI] [PubMed] [Google Scholar]
  3. Berthon B., Binet A., Mauger J. P., Claret M. Cytosolic free Ca2+ in isolated rat hepatocytes as measured by quin2. Effects of noradrenaline and vasopressin. FEBS Lett. 1984 Feb 13;167(1):19–24. doi: 10.1016/0014-5793(84)80824-8. [DOI] [PubMed] [Google Scholar]
  4. Burgess G. M., McKinney J. S., Fabiato A., Leslie B. A., Putney J. W., Jr Calcium pools in saponin-permeabilized guinea pig hepatocytes. J Biol Chem. 1983 Dec 25;258(24):15336–15345. [PubMed] [Google Scholar]
  5. Chan K. M., Junger K. D. Calcium transport and phosphorylated intermediate of (Ca2+ + Mg2+)-ATPase in plasma membranes of rat liver. J Biol Chem. 1983 Apr 10;258(7):4404–4410. [PubMed] [Google Scholar]
  6. Charest R., Blackmore P. F., Berthon B., Exton J. H. Changes in free cytosolic Ca2+ in hepatocytes following alpha 1-adrenergic stimulation. Studies on Quin-2-loaded hepatocytes. J Biol Chem. 1983 Jul 25;258(14):8769–8773. [PubMed] [Google Scholar]
  7. Cockcroft S., Taylor J. A. Fluoroaluminates mimic guanosine 5'-[gamma-thio]triphosphate in activating the polyphosphoinositide phosphodiesterase of hepatocyte membranes. Role for the guanine nucleotide regulatory protein Gp in signal transduction. Biochem J. 1987 Jan 15;241(2):409–414. doi: 10.1042/bj2410409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Combettes L., Berthon B., Binet A., Claret M. Glucagon and vasopressin interactions on Ca2+ movements in isolated hepatocytes. Biochem J. 1986 Aug 1;237(3):675–683. doi: 10.1042/bj2370675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dawson A. P., Comerford J. G., Fulton D. V. The effect of GTP on inositol 1,4,5-trisphosphate-stimulated Ca2+ efflux from a rat liver microsomal fraction. Is a GTP-dependent protein phosphorylation involved? Biochem J. 1986 Mar 1;234(2):311–315. doi: 10.1042/bj2340311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dawson A. P., Irvine R. F. Inositol (1,4,5)trisphosphate-promoted Ca2+ release from microsomal fractions of rat liver. Biochem Biophys Res Commun. 1984 May 16;120(3):858–864. doi: 10.1016/s0006-291x(84)80186-2. [DOI] [PubMed] [Google Scholar]
  11. Downes C. P., Michell R. H. The polyphosphoinositide phosphodiesterase of erythrocyte membranes. Biochem J. 1981 Jul 15;198(1):133–140. doi: 10.1042/bj1980133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Epping R. J., Bygrave F. L. A procedure for the rapid isolation from rat liver of plasma membrane vesicles exhibiting Ca2+-transport and Ca2+-ATPase activities. Biochem J. 1984 Nov 1;223(3):733–745. doi: 10.1042/bj2230733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Evans W. H. A biochemical dissection of the functional polarity of the plasma membrane of the hepatocyte. Biochim Biophys Acta. 1980 May 27;604(1):27–64. doi: 10.1016/0005-2736(80)90584-2. [DOI] [PubMed] [Google Scholar]
  14. Fiskum G. Intracellular levels and distribution of Ca2+ in digitonin-permeabilized cells. Cell Calcium. 1985 Apr;6(1-2):25–37. doi: 10.1016/0143-4160(85)90032-6. [DOI] [PubMed] [Google Scholar]
  15. Goodman F. R., Weiss G. B., Adams H. R. Alterations by neomycin of 45Ca movements and contractile responses in vascular smooth muscle. J Pharmacol Exp Ther. 1974 Feb;188(2):472–480. [PubMed] [Google Scholar]
  16. Guillemette G., Balla T., Baukal A. J., Catt K. J. Characterization of inositol 1,4,5-trisphosphate receptors and calcium mobilization in a hepatic plasma membrane fraction. J Biol Chem. 1988 Apr 5;263(10):4541–4548. [PubMed] [Google Scholar]
  17. Irvine R. F., Moor R. M. Micro-injection of inositol 1,3,4,5-tetrakisphosphate activates sea urchin eggs by a mechanism dependent on external Ca2+. Biochem J. 1986 Dec 15;240(3):917–920. doi: 10.1042/bj2400917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kraus-Friedmann N., Biber J., Murer H., Carafoli E. Calcium uptake in isolated hepatic plasma-membrane vesicles. Eur J Biochem. 1982 Dec;129(1):7–12. doi: 10.1111/j.1432-1033.1982.tb07014.x. [DOI] [PubMed] [Google Scholar]
  19. Kuno M., Gardner P. Ion channels activated by inositol 1,4,5-trisphosphate in plasma membrane of human T-lymphocytes. Nature. 1987 Mar 19;326(6110):301–304. doi: 10.1038/326301a0. [DOI] [PubMed] [Google Scholar]
  20. Lipsky J. J., Lietman P. S. Aminoglycoside inhibition of a renal phosphatidylinositol phospholipase C. J Pharmacol Exp Ther. 1982 Feb;220(2):287–292. [PubMed] [Google Scholar]
  21. Lotersztajn S., Hanoune J., Pecker F. A high affinity calcium-stimulated magnesium-dependent ATPase in rat liver plasma membranes. Dependence of an endogenous protein activator distinct from calmodulin. J Biol Chem. 1981 Nov 10;256(21):11209–11215. [PubMed] [Google Scholar]
  22. Marche P., Koutouzov S., Girard A. Impairment of membrane phosphoinositide metabolism by aminoglycoside antibiotics: streptomycin, amikacin, kanamycin, dibekacin, gentamicin and neomycin. J Pharmacol Exp Ther. 1983 Nov;227(2):415–420. [PubMed] [Google Scholar]
  23. Mauger J. P., Poggioli J., Claret M. Synergistic stimulation of the Ca2+ influx in rat hepatocytes by glucagon and the Ca2+-linked hormones vasopressin and angiotensin II. J Biol Chem. 1985 Sep 25;260(21):11635–11642. [PubMed] [Google Scholar]
  24. Mauger J. P., Poggioli J., Guesdon F., Claret M. Noradrenaline, vasopressin and angiotensin increase Ca2+ influx by opening a common pool of Ca2+ channels in isolated rat liver cells. Biochem J. 1984 Jul 1;221(1):121–127. doi: 10.1042/bj2210121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Moore L., Chen T., Knapp H. R., Jr, Landon E. J. Energy-dependent calcium sequestration activity in rat liver microsomes. J Biol Chem. 1975 Jun 25;250(12):4562–4568. [PubMed] [Google Scholar]
  26. Parker J. C., Barritt G. J. Evidence that lanthanum ions stimulate calcium inflow to isolated hepatocytes. Biochem J. 1981 Oct 15;200(1):109–114. doi: 10.1042/bj2000109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Poggioli J., Mauger J. P., Claret M. Effect of cyclic AMP-dependent hormones and Ca2+-mobilizing hormones on the Ca2+ influx and polyphosphoinositide metabolism in isolated rat hepatocytes. Biochem J. 1986 May 1;235(3):663–669. doi: 10.1042/bj2350663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Poggioli J., Mauger J. P., Guesdon F., Claret M. A regulatory calcium-binding site for calcium channel in isolated rat hepatocytes. J Biol Chem. 1985 Mar 25;260(6):3289–3294. [PubMed] [Google Scholar]
  29. Prpić V., Green K. C., Blackmore P. F., Exton J. H. Vasopressin-, angiotensin II-, and alpha 1-adrenergic-induced inhibition of Ca2+ transport by rat liver plasma membrane vesicles. J Biol Chem. 1984 Feb 10;259(3):1382–1385. [PubMed] [Google Scholar]
  30. Sastrasinh M., Knauss T. C., Weinberg J. M., Humes H. D. Identification of the aminoglycoside binding site in rat renal brush border membranes. J Pharmacol Exp Ther. 1982 Aug;222(2):350–358. [PubMed] [Google Scholar]
  31. Schanne F. A., Moore L. Liver plasma membrane calcium transport. Evidence for a Na+-dependent Ca2+ flux. J Biol Chem. 1986 Jul 25;261(21):9886–9889. [PubMed] [Google Scholar]
  32. Tsien R. Y., Pozzan T., Rink T. J. Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator. J Cell Biol. 1982 Aug;94(2):325–334. doi: 10.1083/jcb.94.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Uhing R. J., Prpic V., Jiang H., Exton J. H. Hormone-stimulated polyphosphoinositide breakdown in rat liver plasma membranes. Roles of guanine nucleotides and calcium. J Biol Chem. 1986 Feb 15;261(5):2140–2146. [PubMed] [Google Scholar]
  34. Whipps D. E., Armston A. E., Pryor H. J., Halestrap A. P. Effects of glucagon and Ca2+ on the metabolism of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate in isolated rat hepatocytes and plasma membranes. Biochem J. 1987 Feb 1;241(3):835–845. doi: 10.1042/bj2410835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. von Tscharner V., Prod'hom B., Baggiolini M., Reuter H. Ion channels in human neutrophils activated by a rise in free cytosolic calcium concentration. 1986 Nov 27-Dec 3Nature. 324(6095):369–372. doi: 10.1038/324369a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES