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. 1998 Aug;18(4):379–390. doi: 10.1023/A:1022593330685

Protein Kinase C Controls the Priming Step of Regulated Exocytosis in Adrenal Chromaffin Cells

Hiroaki Misonou 1, Mica Ohara-Imaizumi 1, Takeshi Murakami 1, Masakazu Kawasaki 1, Ken Ikeda 1, Takeshi Wakai 1, Konosuke Kumakura 1
PMCID: PMC11560201  PMID: 9619293

Abstract

1. To investigate the mechanism whereby protein kinase C enhances secretory function in adrenal chromaffin cells, we examined the effects of 12-O-tetradecanoylphorbor-13-acetate (TPA) on Ca2+-induced catecholamine release from digitonin-permeabilized cells, resolving the release into a MgATP-dependent priming step and a MgATP-indepen-dent Ca2+-triggered step. Treatment with TPA selectively potentiated the priming activityof MgATP, with little increase in the MgATP-independent release. The potentiation by TPA of the MgATP-dependent priming was blocked by [Ser25]protein kinase C(19-31),a specific substrate of protein kinase C. Gö 6976, an inhibitor selective for protein kinase C α and β isoforms, also blocked the potentiation by TPA. These results suggest that activation of protein kinase C, probably the α isoform, potentiates the MgATP-dependent priming step.

2. The antibody raised against GAP-43, a known substrate of protein kinase C, also potentiated the MgATP-dependent priming. The effect of TPA and that of the anti-GAP-43 antibody were not additive. Calmodulin, which binds to GAP-43 and inhibits its phosphorylation by protein kinase C, abolished the effect of TPA. Thus, the present results suggest that protein kinase C potentiates MgATP-dependent priming, at least in part, through phosphorylation of GAP-43.

Keywords: protein kinase C, phorbor ester, GAP-43, priming of exocytosis, secretion

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