title: Protein kinase C activation has distinct effects on the localization, phosphorylation and detergent solubility of the claudin protein family in tight and leaky epithelial cells.
abstract: We have previously shown that protein kinase C (PKC) activation has distinct effects on the structure and barrier properties of cultured epithelial cells (HT29 and MDCK I). Since the claudin family of tight junction (TJ)-associated proteins is considered to be crucial for the function of mature TJ, we assessed their expression patterns and cellular destination, detergent solubility and phosphorylation upon PKC stimulation for 2 or 18 h with phorbol myristate acetate (PMA). In HT29 cells, claudins 1, 3, 4 and 5 and possibly claudin 2 were redistributed to apical cell-cell contacts after PKC activation and the amounts of claudins 1, 3 and 5, but not of claudin 2, were increased in cell lysates. By contrast, in MDCK I cells, PMA treatment resulted in redistribution of claudins 1, 3, 4 and 5 from the TJ and in reorganization of the proteins into more insoluble complexes. Claudins 1 and 4 were phosphorylated in both MDCK I and HT29 cells, but PKC-induced changes in claudin phosphorylation state were detected only in MDCK I cells. A major difference between HT29 and MDCK I cells, which have low and high basal transepithelial electrical resistance, respectively, was the absence of claudin 2 in the latter. Our findings show that PKC activation targets in characteristic ways the expression patterns, destination, detergent solubility and phosphorylation state of claudins in epithelial cells with different capacities to form an epithelial barrier.
authors.affiliation: Division of Medical Microbiology, Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden. anita.sjo@liu.se.
authors.forename: Anita
authors.initials: A
authors.lastname: Sjö
authors.forename: Karl-Eric
authors.initials: KE
authors.lastname: Magnusson
authors.forename: Kajsa Holmgren
authors.initials: KH
authors.lastname: Peterson
mesh_terms: D000818:Animals; D002460:Cell Line; D062445:Claudin-1; D057167:Claudins / Q000378:metabolism*; D004285:Dogs; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D019073:HT29 Cells; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D008565:Membrane Proteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
chemical_list: C568108:CLDN1 protein, human; D062445:Claudin-1; D057167:Claudins; D008565:Membrane Proteins; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate