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Aquaporin-2 Ser-261 phosphorylation is regulated in combination with Ser-256 and Ser-269 phosphorylation
- Source :
- Biochemical and Biophysical Research Communications. 482:524-529
- Publication Year :
- 2017
- Publisher :
- Elsevier BV, 2017.
-
Abstract
- Aquaporin-2 (AQP2) is a water channel in collecting duct principal cells in the kidney. Vasopressin catalyzes AQP2 phosphorylation at several serine sites in its C-terminus: Ser-256, Ser-261, and Ser-269. Upon stimulation by vasopressin, Ser-269 phosphorylation increases and Ser-261 phosphorylation decreases. Ser-256 phosphorylation is relatively constant. However, whether these types of phospho-regulation occur independently in distinct AQP2 populations or sequentially in the same AQP2 population is unclear. Especially, the manner of vasopressin-mediated Ser-261 phospho-regulation has been in controversy. In this study, we established phospho-specific AQP2 immunoprecipitation assays and investigated how pS256-positive AQP2 and pS269-positive AQP2 are catalyzed by forskolin or vasopressin, focusing on their Ser-261 phosphorylation status in polarized Madin-Darby canine kidney (MDCK) cells and in mice. In forskolin-treated MDCK cells, Ser-269 phosphorylation preceded Ser-261 dephosphorylation and Ser-256 phosphorylation was constant. In both MDCK cells and mouse kidney, phospho-specific immunoprecipitation revealed that the regulated Ser-269 phosphorylation occurred in the pS256-positive AQP2 population. Importantly, basal-state Ser-261 phosphorylation and its regulated dephosphorylation occurred in the pS256- and pS269-positive AQP2 population. These results provide the direct evidence that the Ser-261 dephosphorylation is involved in the pS256- and pS269-related AQP2 regulation.
- Subjects :
- 0301 basic medicine
Vasopressin
Vasopressins
Immunoprecipitation
Population
030232 urology & nephrology
Biophysics
macromolecular substances
Biology
Kidney
urologic and male genital diseases
Biochemistry
Cell Line
Dephosphorylation
Mice
03 medical and health sciences
chemistry.chemical_compound
Dogs
0302 clinical medicine
Serine
Animals
Protein phosphorylation
Phosphorylation
education
Molecular Biology
education.field_of_study
Aquaporin 2
Forskolin
urogenital system
Colforsin
Cell Biology
Molecular biology
Rats
Mice, Inbred C57BL
030104 developmental biology
chemistry
Subjects
Details
- ISSN :
- 0006291X
- Volume :
- 482
- Database :
- OpenAIRE
- Journal :
- Biochemical and Biophysical Research Communications
- Accession number :
- edsair.doi.dedup.....c948dcde7db92e1b31cfcc0e7c33f941