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J Biol Chem. 1997 Jun 20;272(25):16056-61.

Heterotrimeric G-protein Gq/11 localized on pancreatic zymogen granules is involved in calcium-regulated amylase secretion.

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  • 1Department of Physiology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.

Abstract

The heterotrimeric G-protein Gq/11 was identified on pancreatic acinar zymogen granules and its function in calcium-regulated exocytosis was examined. Western blotting showed alphaq/11, but not alphas or alphao, to be localized to the zymogen granule membrane along with G-protein beta-subunit; all three alpha subunits were present in a plasma membrane fraction and the alphaq/11 signal was 30-fold more enriched in the plasma membrane as compared with granule membrane. Neither CCK receptors nor alpha subunits of the sodium pump, both plasma membrane markers were present on granule membranes. Immunohistochemistry of pancreatic lobules showed that alphaq/11 localized to the zymogen granule-rich apical region of acinar cells together with a much stronger signal at the basolateral plasma membrane. When the substance-P-related peptide GPAnt-2a, an antagonist of Gq/11, was introduced into streptolysin-O permeabilized acini to bypass the plasma membrane, the amylase release induced by 10 microM free calcium was potentiated in a concentration-dependent manner. By contrast, another substance-P-related peptide, GPAnt-1, an antagonist of Go and Gi, showed no effect on calcium-induced amylase release from permeabilized acini. GPAnt-2a peptide also exerted an inhibitory effect on the total GTPase activity of the purified zymogen granules and a larger inhibitory effect on the GTPase activity of the Gq/11 protein immunopurified from zymogen granules. GPAnt-1, however, did not inhibit GTPase activity of either zymogen granules or immunopurified Gq/11. These results suggest that GPAnt-2a peptide augmented calcium-induced amylase release from permeabilized acini by inhibiting GTPase activity of the Gq/11 protein on zymogen granules. We conclude that Gq/11 protein on zymogen granules plays a tonic inhibitory role in calcium-regulated amylase secretion from pancreatic acini.

PMID:
9188511
[PubMed - indexed for MEDLINE]
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