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Protein kinase Cδ mediates cyclic adenosine monophosphate–stimulated translocation of sodium taurocholate cotransporting polypeptide and multidrug resistant associated protein 2 in rat hepatocytes
Author(s) -
Schonhoff Christopher M.,
Gillin Henry,
Webster Cynthia R. L.,
Anwer M. Sawkat
Publication year - 2008
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.22162
Subject(s) - cyclic adenosine monophosphate , adenosine , protein kinase a , chemistry , chromosomal translocation , kinase , microbiology and biotechnology , biochemistry , biology , receptor , gene
Cyclic adenosine monophosphate (cAMP) stimulates translocation of Na + ‐taurocholate (TC) cotransporting polypeptide (Ntcp) and multidrug resistant associated protein 2 (Mrp2) to the plasma membrane. Because cAMP activates phosphoinositide‐3‐kinase (PI3K) and protein kinase C (PKC) activation is PI3K‐dependent, the aim of the current study was to determine whether cAMP activates conventional and novel PKCs in hepatocytes and whether such activation plays a role in cAMP‐stimulated Ntcp and Mrp2 translocation. The effect of cAMP on PKCs, TC uptake, and Ntcp and Mrp2 translocation was studied in isolated rat hepatocytes using a cell‐permeable cAMP analog, CPT‐cAMP. The activity of PKCs was assessed from membrane translocation of individual PKCs, and phospho‐specific antibodies were used to determine PKCδ phosphorylation. TC uptake was determined from time‐dependent uptake of 14 C‐TC, and a cell surface biotinylation method was used to determine Ntcp and Mrp2 translocation. CPT‐cAMP stimulated nPKCδ but not cPKCα or nPKCϵ, and induced PI3K‐dependent phosphorylation of nPKCδ at Thr 505 . Rottlerin, an inhibitor of nPKCδ, inhibited cAMP‐induced nPKCδ translocation, TC uptake, and Ntcp and Mrp2 translocation. Bistratene A, an activator of nPKCδ, stimulated nPKCδ translocation, TC uptake, and Ntcp and Mrp2 translocation. The effects of cAMP and bistratene A on TC uptake and Ntcp and Mrp2 translocation were not additive. Conclusion: These results suggest that cAMP stimulates Ntcp and Mrp2 translocation, at least in part, by activating nPKCδ via PI3K‐dependent phosphorylation at Thr 505 . (H EPATOLOGY 2008.)