Protein Kinase A Associates with Cystic Fibrosis Transmembrane Conductance Regulator via an Interaction with Ezrin *
Journal of Biological Chemistry, ISSN: 0021-9258, Vol: 275, Issue: 19, Page: 14360-14366
2000
- 143Citations
- 45Captures
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Metrics Details
- Citations143
- Citation Indexes143
- 143
- CrossRef119
- Captures45
- Readers45
- 45
Article Description
The cystic fibrosis transmembrane conductance regulator (CFTR) is an epithelial Cl − channel whose activity is controlled by cAMP-dependent protein kinase (PKA)-mediated phosphorylation. We found that CFTR immunoprecipitates from Calu-3 airway cells contain endogenous PKA, which is capable of phosphorylating CFTR. This phosphorylation is stimulated by cAMP and inhibited by the PKA inhibitory peptide. The endogenous PKA that co-precipitates with CFTR could also phosphorylate the PKA substrate peptide, Leu-Arg-Arg-Ala-Ser-Leu-Gly (kemptide). Both the catalytic and type II regulatory subunits of PKA are identified by immunoblotting CFTR immunoprecipitates, demonstrating that the endogenous kinase associated with CFTR is PKA, type II (PKA II). Phosphorylation reactions mediated by CFTR-associated PKA II are inhibited by Ht31 peptide but not by the control peptide Ht31P, indicating that a protein kinase A anchoring protein (AKAP) is responsible for the association between PKA and CFTR. Ezrin may function as this AKAP, since it is expressed in Calu-3 and T84 epithelia, ezrin binds RII in overlay assays, and RII is immunoprecipitated with ezrin from Calu-3 cells. Whole-cell patch clamp of Calu-3 cells shows that Ht31 peptide reduces cAMP-stimulated CFTR Cl − current, but Ht31P does not. Taken together, these data demonstrate that PKA II is linked physically and functionally to CFTR by an AKAP interaction, and they suggest that ezrin serves as an AKAP for PKA-mediated phosphorylation of CFTR.
Bibliographic Details
http://www.sciencedirect.com/science/article/pii/S0021925819806087; http://dx.doi.org/10.1074/jbc.275.19.14360; http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0034640510&origin=inward; http://www.ncbi.nlm.nih.gov/pubmed/10799517; https://linkinghub.elsevier.com/retrieve/pii/S0021925819806087; https://dx.doi.org/10.1074/jbc.275.19.14360
Elsevier BV
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