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PMID: 3021780 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Isolated secretion granules from parotid glands of chronically stimulated rats possess an alkaline internal pH and inward-directed H+ pump activity.

The Journal of cell biology ·Vol. 103 ·No. 4 ·1986-10-00 ·Pages 1257-67

Arvan P, Castle JD

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.

MeSH Terms
Adenosine Triphosphatases/metabolism Adenosine Triphosphate/metabolism Animals Cytoplasmic Granules/metabolism H(+)-K(+)-Exchanging ATPase Hydrogen-Ion Concentration Isoproterenol/pharmacology Male Membrane Potentials/drug effects Microscopy, Fluorescence Parotid Gland/drug effects,metabolism Rats Rats, Inbred Strains Salivary Proteins and Peptides/metabolism Stimulation, Chemical
Chemicals
Salivary Proteins and Peptides Adenosine Triphosphate Adenosine Triphosphatases H(+)-K(+)-Exchanging ATPase Isoproterenol
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Arvan P
Castle J D
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44 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.103.4.1257
Published
1986-10-00
Pages
1257-67
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2114332
Subset
IM
Grants
NIGMS NIH HHS · GM 07205 · United States
NIGMS NIH HHS · GM 26524 · United States
External Links
PubMed source DOI: 10.1083/jcb.103.4.1257 PMC Full Text View journal details
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