N.A.S. Ismail
The phosphorylation of endogenous Nedd4-2 In Na+—absorbing human airway epithelial cells
Ismail, N.A.S.; Baines, D.L.; Wilson, S.M.
Authors
D.L. Baines
S.M. Wilson
Abstract
Neural precursor cell expressed, developmentally down-regulated protein 4-2 (Nedd4-2) mediates the internalisation / degradation of epithelial Na+ channel subunits (α-, β- and γ-ENaC). Serum / glucocorticoid inducible kinase 1 (SGK1) and protein kinase A (PKA) both appear to inhibit this process by phosphorylating Nedd4-2-Ser221, -Ser327 and -Thr246. This Nedd4-2 inactivation process is thought to be central to the hormonal control of Na+ absorption. The present study of H441 human airway epithelial cells therefore explores the effects of SGK1 and / or PKA upon the phosphorylation / abundance of endogenous Nedd4-2; the surface expression of ENaC subunits, and electrogenic Na+ transport. Effects on Nedd4-2 phosphorylation/abundance and the surface expression of ENaC were monitored by western analysis, whilst Na+ absorption was quantified electrometrically. Acutely (20 min) activating PKA in glucocorticoid-deprived (24 h) cells increased the abundance of Ser221-phosphorylated, Ser327-phosphorylated and total Nedd4-2 without altering the abundance of Thr246-phosphorylated Nedd4-2. Activating PKA under these conditions did not cause a co-ordinated increase in the surface abundance of α-, β- and γ-ENaC and had only a very small effect upon electrogenic Na+ absorption. Activating PKA (20 min) in glucocorticoid-treated (0.2 µM dexamethasone, 24 h) cells, on the other hand, increased the abundance of Ser221-, Ser327- and Thr246-phosphorylated and total Nedd4-2; increased the surface abundance of α-, β- and γ-ENaC and evoked a clear stimulation of Na+ transport. Chronic glucocorticoid stimulation therefore appears to allow cAMP-dependent control of Na+ absorption by facilitating the effects of PKA upon the Nedd4-2 and ENaC subunits.
Citation
Ismail, N., Baines, D., & Wilson, S. (2014). The phosphorylation of endogenous Nedd4-2 In Na+—absorbing human airway epithelial cells. European Journal of Pharmacology, 732, 32-42. https://doi.org/10.1016/j.ejphar.2014.03.005
Journal Article Type | Article |
---|---|
Publication Date | Jun 5, 2014 |
Deposit Date | Feb 27, 2014 |
Publicly Available Date | Jun 17, 2014 |
Journal | European Journal of Pharmacology |
Print ISSN | 0014-2999 |
Publisher | Elsevier |
Peer Reviewed | Peer Reviewed |
Volume | 732 |
Pages | 32-42 |
DOI | https://doi.org/10.1016/j.ejphar.2014.03.005 |
Keywords | Epithelial Na+ channel, Serum and glucocorticoid regulated kinase 1, Protein kinase A, Pulmonary Na+ absorption, Cellular signalling. |
Files
Published Journal Article
(2.2 Mb)
PDF
Publisher Licence URL
http://creativecommons.org/licenses/by/3.0/
Copyright Statement
© 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license
(http://creativecommons.org/licenses/by/3.0/)
You might also like
Downloadable Citations
About Durham Research Online (DRO)
Administrator e-mail: dro.admin@durham.ac.uk
This application uses the following open-source libraries:
SheetJS Community Edition
Apache License Version 2.0 (http://www.apache.org/licenses/)
PDF.js
Apache License Version 2.0 (http://www.apache.org/licenses/)
Font Awesome
SIL OFL 1.1 (http://scripts.sil.org/OFL)
MIT License (http://opensource.org/licenses/mit-license.html)
CC BY 3.0 ( http://creativecommons.org/licenses/by/3.0/)
Powered by Worktribe © 2024
Advanced Search