Protease-activated receptor-2 increases exocytosis via multiple signal transduction pathways in pancreatic duct epithelial cells
SCIE
SCOPUS
- Title
- Protease-activated receptor-2 increases exocytosis via multiple signal transduction pathways in pancreatic duct epithelial cells
- Authors
- Kim, MH; Choi, BH; Jung, SR; Sernka, TJ; Kim, S; Kim, KT; Hille, B; Nguyen, TD; Koh, DS
- Date Issued
- 2008-07-04
- Publisher
- AMER SOC BIOCHEMISTRY MOLECULAR BIOLO
- Abstract
- Protease-activated receptor-2 (PAR-2) is activated when trypsin cleaves its NH2 terminus to expose a tethered ligand. We previously demonstrated that PAR-2 activates ion channels in pancreatic duct epithelial cells (PDEC). Using real-time optical fluorescent probes, cyan fluorescence protein-Epac1-yellow fluorescence protein for cAMP, PHPLC-delta 1-enhanced green fluorescent protein for phosphatidylinositol 4,5-bisphosphate, and protein kinase C gamma (PKC gamma)-C1-yellow fluorescence protein for diacylglycerol, we now define the signaling pathways mediating PAR-2 effect in dog PDEC. Although PAR-2 activation does not stimulate acAMPincrease, it induces phospholipase C to hydrolyze phosphatidylinositol 4,5-bisphosphate into inositol 1,4,5-trisphosphate and diacylglycerol. Intracellular Ca2+ mobilization from inositol 1,4,5-trisphosphate- sensitive Ca2+ stores and a subsequent Ca2+ influx through store-operated Ca2+ channels cause a biphasic increase in intracellular Ca2+ concentration ([Ca2+](i)), measured with Indo-1 dye. Single-cell amperometry demonstrated that this increase in [Ca2+] i in turn causes a biphasic increase in exocytosis. A protein kinase assay revealed that trypsin also activates PKC isozymes to stimulate additional exocytosis. Paralleling the increased exocytosis, mucin secretion from PDEC was also induced by trypsin or the PAR-2 activating peptide. Consistent with the serosal localization of PAR-2, 1 mu M luminal trypsin did not induce exocytosis in polarized PDEC monolayers; on the other hand, 10 mu M trypsin at 37 degrees C damaged the epithelial barrier sufficiently so that it could reach and activate the serosal PAR-2 to stimulate exocytosis. Thus, in PDEC, PAR-2 activation increases [Ca2+](i) and activates PKC to stimulate exocytosis and mucin secretion. These functions may mediate the reported protective role of PAR-2 in different models of pancreatitis.
- Keywords
- MOLECULAR-CLONING; SECRETION; AMPEROMETRY; CA2+; MECHANISMS; PROTECTION
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/22683
- DOI
- 10.1074/jbc.M801655200
- ISSN
- 0021-9258
- Article Type
- Article
- Citation
- JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 283, no. 27, page. 18711 - 18720, 2008-07-04
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