Protein kinase C-alpha attenuates cholinergically stimulated gastric acid secretion of rabbit parietal cells.
(1) The phorbolester 12-O-tetradecanoyl phorbol-13-acetate (TPA), an activator of protein kinase C (PKC), inhibits cholinergic stimulation of gastric acid secretion. We observed that this effect strongly correlated with the inhibition of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) activity in rabbit parietal cells. (2) The aim of this study was to specify the function of PKC-alpha in cholinergically stimulated H(+) secretion. PKC-alpha represents the only calcium-dependent PKC isoenzyme that has been detected in rabbit parietal cells. (3) Gö 6976, an inhibitor of calcium-dependent PKC, concentration-dependently antagonized the inhibitory effect of TPA, and, therefore, revealed the action of PKC-alpha on carbachol-induced acid secretion in rabbit parietal cells. (4) TPA exerted no additive inhibition of carbachol-stimulated acid secretion if acid secretion was partially inhibited by the potent CaMKII inhibitor 1-[N,O-bis(5-isoquinolinsulfonyl)-N-methyl-L-tyrosyl]-4-phenyl-piperazine (KN-62). (5) Since both kinase modulators, TPA and KN-62, affected no divergent signal transduction pathways in the parietal cell, an in vitro model has been used to study if PKC directly targets CaMKII. CaMKII purified from parietal cell-containing gastric mucosa of pig, was transphosphorylated by purified cPKC containing PKC-alpha up to 1.8 mol P(i) per mol CaMKII in vitro. The autonomy site of CaMKII was not transphosphorylated by PKC. (6) The phosphotransferase activity of the purified CaMKII was in vitro inhibited after transphosphorylation by PKC if calmodulin was absent during transphosphorylation. Attenuation of CaMKII activity by PKC showed strong similarity to the downregulation of CaMKII by basal autophosphorylation. (7) Our results suggest that PKC-alpha and CaMKII are closely functionally linked in a cholinergically induced signalling pathway in rabbit parietal cells. We assume that in cholinergically stimulated parietal cells PKC-alpha transinhibits CaMKII activity, resulting in an attenuation of acid secretion.
Fährmann
Michael
M
Institut für Zoophysiologie der Westfälischen Wilhelms-Universität Münster, Hindenburgplatz 55, D-48143 Münster, Germany. Faehrmann.michael@mh-hannover.de
Kaufhold
Marc
M
Pfeiffer
Andreas F
AF
Seidler
Ursula
U
D000818:Animals; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005744:Gastric Acid / Q000378:metabolism*; D010295:Parietal Cells, Gastric / Q000187:drug effects* / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D051571:Protein Kinase C-alpha; D011817:Rabbits
D004791:Enzyme Inhibitors; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases