45Ca2+ uptake in isolated rat parotid secretory granules was examined in the presence of oxalate. Uptake of calcium was dependent on time, with the maximum occurring at 15 min. The uptake of calcium was dependent on adenosine-5'-triphosphate (ATP), and substitution of ATP with B,g-methylene-ATP did not stimulate calcium uptake. Enzyme marker analysis indicated that mitochondria accounted for no greater than 3.0 ±0.2% of the observed ATP-dependent calcium uptake. Calcium uptake was blocked by the ATPase inhibitors tributyltin, IC 50 = 12.2 ±0.6 nmol/L and 4-acetamido-4'-isothiocyano-2,2'-stilbene disulphonic acid (SITS), IC50 = 3.0 ±0.3 (μmol/L. These results indicate that in the parotid secretory granule there is a calcium uptake mechanism that is dependent on the hydrolysis of ATP and is suppressed by two inhibitors of granule ATPase.
Get full access to this article
View all access options for this article.
References
1.
Apps, D.K.; Pryde, J.G.; and Sutton, R. (1982): The H+-Translocating Adenosine Triphosphatase of Chromaffin Granule Membranes, Ann N YAcad Sci402:134-145.
2.
Baum, B.J.; Ambudkar, I.S.; and Horn, V.J. (1988): Evidence thatATP-Dependent Ca2* Transport in Rat Parotid Microsomal Membranes Requires Charge Compensation, Biochem J254:649-654.
3.
Bayerdorffer, E.; Eckhardt, L.; Haase, W.; and Schulz, I. (1985): Electrogenic Calcium Transport in Plasma Membrane of Rat Pancreatic Acinar Cells, J Membrane Biol84:45-60.
4.
Beers, M.F.; Carty, S.E.; Johnson, R.G.; and Scarpa, A. (1982): H+-ATPase and Catecholamine Transport in Chromaffin Granules, Ann N YAcad Sci402:116-133.
5.
Bernfeld, P. (1955): Amylases, a and ß. In: Methods in Enzymology, Vol. 1, S.P. Colowick and N.O. Kaplan, Eds., NewYork: Academic Press, pp. 149-158.
6.
De Lisle, R.C. and Williams, J.A. (1986): Regulation of Membrane Fusion in Secretory Exocytosis . In: Ann Rev Physiol, Vol. 48, R.M. Berne and J.F. Hoffman, Eds., Palo Alto: Annual Reviews Inc., pp. 225-238.
7.
Dissing, S.; Nauntofte, B.; and Sten-Knudsen, 0. (1990): Spatial Distribution of Intracellular, Free Ca2+ in Isolated Rat Parotid Acini, Pflugers Arch417:1-12.
8.
Dowd, F. (1980): Characterization of a Microsomal Fraction from Rabbit Parotid Gland: Partial Separation of Alkaline Phosphate from Na+-K+-ATPase , Arch Oral Biol25:773-780.
9.
Dowd, F.J.; Pasieniuk, J.A.; Hand, A.R.; Cheung, P.H.; Haines, D.W. (1989): Characteristics of Anion-stimulated Mg-ATPase from Rat Parotid Gland Secretory Granules, Arch Oral Biol34:167-177.
10.
Dowd, F.J. and Porter, J. (1987): Calcium Binding to Parotid Secretory Granules , J Dent Res66:307, Abst. No. 1603.
11.
Green, D.E.; Mii, S.; and Kohout, P.M. (1955): Studies on the Terminal Electron Transport System . I. Succinic Dehydrogenase, J Biol Chem217:551-567.
12.
Grinstein, S. and Furuya, W. (1982): A Mg2+-Stimulated ATPase in Platelet Alpha-Granule Membranes: Possible Involvement in Proton Translocation, Arch Biochem Biophys218:502-512.
13.
Grover, A.K.; Xwan, C.Y.; and Daniel, E.E. (1982): Ca2+ Dependence of Calcium Uptake by Rat Myometrium Plasma Membrane-enriched Fraction, Am J Physiol242:C278-C282.
14.
Harper, J.F. (1988): Stimulus-Secretion Coupling: Second Messenger-Regulated Exocytosis. In: Advances in Second Messenger and Phosphoprotein Research, Vol. 22, P. Greengard and G.A. Robison, Eds., New York: Raven Press, pp.193-318.
15.
Hutton, J.C. and Peshavaria, M. (1982): Proton Translocating Mg2+ Dependent ATPase Activity in Insulin-Secretory Granules, Biochem J204:161-170.
16.
Iversen, J.M.; Kauffman, D.L.; Keller, P.J.; and Robinovitch, M. (1985): Isolation and Partial Characterization of Two Populations of Secretory Granules from Rat Parotid Glands, Cell Tissue Res240 :441-444.
17.
Kanagasuntheram, P. and Teo, T.S. (1982): Parotid Microsomal Calcium Transport. Subcellular Localization and Characterization, Biochem J208:789-794.
18.
Lorenson, M.Y.; Lee, Y.-C.; and Jacobs, L.S. (1981): Identification and Characterization of an Anion-sensitive Mg2+-ATPase in Pituitary Secretory Granule Membranes, J Biol Chem256:12,802-12,810.
19.
Lowry, O.H.; Rosebrough, N.J.; Farr, A.L.; and Randall, R.J. (1951): Protein Measurement with the Folin Phenol Reagent , J Biol Chem193:265-275.
20.
Lucy, J.A. and Ahkong, Q.F. (1986): An Osmotic Model for the Fusion of Biological Membranes, FEBS Lett199:1-11.
21.
Mangos, J.A.; Garrish, M.T.; Wells, R.; Farnham, W.; and Bouchlas, G. (1978): A Micropuncture Study of the Handling of Calcium by the Rat Parotid, J Dent Res57:818-825.
22.
Pazoles, C.J.; Creutz, C.E.; Ramu, A.; and Pollard, H.B. (1980): Permeant Anion Activation of MgATPase Activity in Chromaffin Granules, J Biol Chem255:7863-7869.
23.
Thiyagarajah, P. and Lim, S.C. (1986): The (Ca2+ + Mg2+)-Stimulated ATPase of the Rat Parotid Endoplasmic Reticulum, Biochem J235 :491-498.
24.
Vardanis, A. (1977): Protein Kinase Activity in the Inner Membrane of Mammalian Mitochondria, J Biol Chem252:807-813.
25.
Von Zastrow, M.; Tritton, T.R.; and Castle, J.D. (1984): Identification of L-Ascorbic Acid in Secretory Granules of the Rat Parotid Gland, J Biol Chem259:11,746-11,750.
26.
Wakasugi, H.; Kimura, T.; Haase, W.; Kribben, A.; Kaufmann, R.; and Schulz, I. (1982): Calcium Uptake into Acini from Rat Pancreas : Evidence for Intracellular ATP-dependent Calcium Sequestration, J Membrane Biol65:205-220.
27.
Wallach, D. and Schramm, M. (1971): Calcium and the Exportable Protein in Rat Parotid Gland. Parallel Subcellular Distribution and Concomitant Secretion, Eur J Biochem21:433-437.
28.
Zimmerberg, J.; Sardet, C.; and Epel, D. (1985): Exocytosis of Sea Urchin Egg Cortical Vesicles in vitro is Retarded by Hyperosmotic Sucrose: Xinetics of Fusion Monitored by Quantitative Light-Scattering Microscopy, J Cell Biol101:398-410.
