Isolated Secretion Granules from Parotid Glands of Chronically Stimulated Rats Possess an Alkaline Internal PH and Inward-directed H+ Pump Activity

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 1986 Oct 1

PMID 3021780

Citations 12

Authors

P Arvan

J D Castle

Affiliations

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Abstract

Secretion granules have been isolated from the parotid glands of rats that have been chronically stimulated with the beta-adrenergic agonist, isoproterenol. These granules are of interest because they package a quantitatively different set of secretory proteins in comparison with granules from the normal gland. Polypeptides enriched in proline, glycine, and glutamine, which are known to have pI's greater than 10, replace alpha-amylase (pI's = 6.8) as the principal content species. The internal pH of granules from the treated rats ranges from 7.8 in a potassium sulfate medium to 6.9 in a choline chloride medium. The increased pH over that of normal parotid granules (approximately 6.8) appears to reflect the change in composition of the secretory content. Whereas normal mature parotid granules have practically negligible levels of H+ pumping ATPase activity (Arvan, P., G. Rudnick, and J. D. Castle, 1985, J. Biol. Chem., 260, 14945-14952) the isolated granules from isoproterenol-treated rats undergo a time-dependent internal acidification (approximately 0.2 pH unit) that requires the presence of ATP and is abolished by an H+ ionophore. Additionally, an inside-positive granule transmembrane potential develops after ATP addition that depends upon ATP hydrolysis. Two independent methods have been used that exclude the possibility that contaminating organelles are the source of the H+-ATPase activity. Together these data provide clear evidence for the presence of an H+ pump in the membranes of parotid granules from chronically stimulated rats. However, despite the presence of H+-pump activity, fluorescence microscopy with the weak base, acridine orange, reveals that the intragranular pH in live cells is greater than that of the cytoplasm.

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References

RADLEY J . Ultrastructural changes in the rat submaxillary gland following isoprenaline. Z Zellforsch Mikrosk Anat. 1969; 97(2):196-211. DOI: 10.1007/BF00344757. View

Caro L, VAN Tubergen R, Kolb J . High-resolution autoradiography. I. Methods. J Cell Biol. 1962; 15:173-88. PMC: 2106152. DOI: 10.1083/jcb.15.2.173. View

Hand A . Morphology and cytochemistry of the Golgi apparatus of rat salivary gland acnar cells. Am J Anat. 1971; 130(2):141-57. DOI: 10.1002/aja.1001300203. View

Fairbanks G, Steck T, Wallach D . Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971; 10(13):2606-17. DOI: 10.1021/bi00789a030. View

Castle J, JAMIESON J, Palade G . Radioautographic analysis of the secretory process in the parotid acinar cell of the rabbit. J Cell Biol. 1972; 53(2):290-311. PMC: 2108718. DOI: 10.1083/jcb.53.2.290. View

UDENFRIEND S, Stein S, Bohlen P, Dairman W, Leimgruber W, Weigele M . Fluorescamine: a reagent for assay of amino acids, peptides, proteins, and primary amines in the picomole range. Science. 1972; 178(4063):871-2. DOI: 10.1126/science.178.4063.871. View

Castle J, JAMIESON J, Palade G . Secretion granules of the rabbit parotid gland. Isolation, subfractionation, and characterization of the membrane and content subfractions. J Cell Biol. 1975; 64(1):182-210. PMC: 2109471. DOI: 10.1083/jcb.64.1.182. View

Casey R, Njus D, Radda G, Sehr P . Adenosine triphosphate-evoked catecholamine release in chromatin granules. Osmotic lysis as a consequence of proton translocation. Biochem J. 1976; 158(3):583-8. PMC: 1164013. DOI: 10.1042/bj1580583. View

BARKA T, Burke G . Secretory behaviour of hypertrophic and hyperplastic salivary gland. Histochem J. 1977; 9(4):453-66. DOI: 10.1007/BF01002976. View

10.

Herzog V, Farquhar M . Luminal membrane retrieved after exocytosis reaches most golgi cisternae in secretory cells. Proc Natl Acad Sci U S A. 1977; 74(11):5073-7. PMC: 432101. DOI: 10.1073/pnas.74.11.5073. View

11.

Johnson R, Scarpa A . Protonmotive force and catecholamine transport in isolated chromaffin granules. J Biol Chem. 1979; 254(10):3750-60. View

12.

Muenzer J, Bildstein C, Gleason M, CARLSON D . Purification of proline-rich proteins from parotid glands of isoproterenol-treated rats. J Biol Chem. 1979; 254(13):5623-8. View

13.

Holz R . Measurement of membrane potential of chromaffin granules by the accumulation of triphenylmethylphosphonium cation. J Biol Chem. 1979; 254(14):6703-9. View

14.

Rottenberg H . The measurement of membrane potential and deltapH in cells, organelles, and vesicles. Methods Enzymol. 1979; 55:547-69. DOI: 10.1016/0076-6879(79)55066-6. View

15.

BLOOM G, Carlsoo B, Danielsson A, Gustafsson H, Henriksson R . Quantitative structural analysis and the secretory behaviour of the rat parotid gland after long and short term isoprenaline treatment. Med Biol. 1979; 57(4):224-33. View

16.

Abe K, Dawes C . The secretion of protein and of some electrolytes in response to alpha- and beta- adrenergic agonists by rat parotid and submandibular salivary glands enlarged by chronic treatment with isoproterenol. J Dent Res. 1980; 59(6):1081-9. DOI: 10.1177/00220345800590062001. View

17.

Johnson R, Carty S, Fingerhood B, Scarpa A . The internal pH of mast cell granules. FEBS Lett. 1980; 120(1):75-9. DOI: 10.1016/0014-5793(80)81050-7. View

18.

Carty S, Johnson R, Scarpa A . The isolation of intact adrenal chromaffin granules using isotonic Percoll density gradients. Anal Biochem. 1980; 106(2):438-45. DOI: 10.1016/0003-2697(80)90545-x. View

19.

Fletcher D, Quigley J, BAUER G, Noe B . Characterization of proinsulin- and proglucagon-converting activities in isolated islet secretory granules. J Cell Biol. 1981; 90(2):312-22. PMC: 2111882. DOI: 10.1083/jcb.90.2.312. View

20.

Loh Y, Gainer H . Characterization of pro-opiocortin-converting activity in purified secretory granules from rat pituitary neurointermediate lobe. Proc Natl Acad Sci U S A. 1982; 79(1):108-12. PMC: 345671. DOI: 10.1073/pnas.79.1.108. View