Involvement of a phosphoprotein on the zymogen granule membrane in the control of regulated exocytosis in the exocrine pancreas.
The pancreatic acinar cell is one of a number of cell types in which phosphoproteins are believed to be involved in the control of regulated exocytosis. We have examined the effects of three agents that affect secretion in the acinar cell on the phosphorylation states of proteins on the zymogen granule membrane. We show that Ca2+ and GTP gamma S, which stimulate secretion, also stimulate the phosphorylation of a protein of M(r) 45,000 (p45) on isolated zymogen granules. On the other hand, the protein kinase inhibitor genistein inhibits both secretion and phosphorylation of p45. For all three agents, p45 phosphorylation is affected over concentration ranges identical to those that affect secretion. The stimulatory effect of GTP gamma S and the inhibitory effect of genistein are also seen when the phosphorylation state of p45 on granules within permeabilized cells is examined. Ca2+, however, over the same concentration range, now causes dephosphorylation of p45. Furthermore, the time-course of this effect is similar to that of Ca(2+)-triggered secretion. Phosphorylation of p45 is exclusively on serine, with no detectable phosphorylation on either threonine or tyrosine. We propose that exocytosis in pancreatic acini is controlled at least in part through the phosphorylation/dephosphorylation of p45, with dephosphorylation acting as a trigger for exocytosis.
MacLean
C M
CM
Department of Pharmacology, University of Cambridge, UK.
Marciniak
S J
SJ
Hall
D V
DV
Edwardson
J M
JM
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000378:metabolism*; D004792:Enzyme Precursors / Q000378:metabolism; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology*; D019833:Genistein; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D066298:In Vitro Techniques; D007529:Isoflavones / Q000494:pharmacology; D008297:Male; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D008970:Molecular Weight; D010179:Pancreas / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D051381:Rats; D017207:Rats, Sprague-Dawley
D004792:Enzyme Precursors; D007529:Isoflavones; D008565:Membrane Proteins; D010750:Phosphoproteins; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D019833:Genistein; D011505:Protein-Tyrosine Kinases; D002118:Calcium